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THE 



Examination of the Urine 



OF THE 



HORSE AND MAN 



PIERRE A. FISH, D.Sc, D.V.M. 

PROFESSOR OF VETERINARY PHYSIOLOGY AND PHARMACOLOGY 

NEW YORK STATE VETERINARY COLLEGE 

CORNELL UNIVERSITY 



PUBLISHED BY 

TAYLOR AND CARPENTER 

ITHACA, N. Y. 

1906 



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COPYRIGHT, I906 

BY 
PIEKRE A. FISH 



PREFACE. 



This manual represents the published form of mimeographed sheets, which 
the writer has used, for some years past, in giving instruction to his veterinary 
students. Data relating to the urine of the horse is not abundant and, unfor- 
tunately, some of the tests which may' be standard for human urine are not 
reliable for that of the horse and, therefore, require modification. In order to 
emphasize this fact, the writer has employed the comparative method and asks 
the students to examine their own urine along with that of the horse and note 
carefully the differences in the results. 

The importance of urine examination for diagnosis and prognosis in human 
medicine is too well known to require emphasis here ; but, notwithstanding 
certain difficulties in the way of its application in veterinary medicine, the writer 
believes that there is no important reason why just as much valuable information 
may not be derived, in certain cases, from the urine of the horse by an up-to- 
date veterinarian, as the physician obtains from the urine of his patients. 

Simplification of methods, without too great a sacrifice in accuracy, is 
essential for satisfactory examination. — especially by veterinarians. The writer 
realizes that only a short step has been taken in this direction, but hopes that 
time will lighten the difficulties. 

P. A. F. 

October, 1906. 



TABLE OF CONTENTS. 



CHAPTER I. 

The Secretion of Urine pp. 5-10 

CHAPTER II. 

Quantity, Color, Transparency, Consistency pp. 10-16 

CHAPTER III. 

Qualitative tests : Water, Chlorides, Sulphates, 

Phosphates, Carbonates pp. 16-20 

CHAPTER IV. 

Organic Constituents: Urea, Uric Acid, Hippuric 
Acid, Creatinin, Mucus, Indican, Oxalic Acid, 
Acetone, Urobilin, Leucin, Tyrosin, Phenol. .. pp. 21-29 
chapter v. 

Abnormal Substances in the Urine: Albumin, Sugar, pp 29-36 

CHAPTER VI. 

Bile, Blood, Melanine pp. 37-40 

CHAPTER VII. 

Quantitative Analysis, Centrifugal Method : Phos- 
phates, Chlorides, Sulphates, Uric Acid, Uri- 
cometer, Urea, Albumen, Saccharometer pp. 40-50 

CHAPTER VIII. 

Volumetric Methods : Chlorides, Phosphoric Acid, 

Sulphuric Acid pp. 50-54 

CHAPTER IX. 

Chemical Examination of Urinary Deposits, Acid 
Urate of Soda, Uric Acid, Cystine, Pus, Blood, 
Alkaline Urates, Triple Phosphates, Calcium 

Oxalate, Calcium Carbonate pp. 55-57 

chapter x. 

Microscopical Examination of Urine, Unorganized 
Sediments : Uric Acid, Acid Urate Soda, Oxa- 
late of Lime, Hippuric Acid, Calcium Sulphate, 
Calcium Phosphate, Triple Phosphate, Urate 
of Ammonium, Cystin, Leucin, Tyrosin. Or- 
ganized Sediments : Epithelial cells; Hyaline, 
Granular, Epithelial, Fat, Hemorrhacic and 
Waxy casts; Cylindroids; Blood and Pus corpus- 
cles ; Spermatozoa ; Mucus ; Bacteria pp. 58-67 



URINE ANALYSIS. 



APPARATUS FOR THE LOCKER. 



t dozen test tubes, 6 inch 

1 dozen test tubes, S inch 

1 Minim pipette 

1 Beaker, 10 oz. 

1 Graduate, 30 cc. 

1 Graduate, 250 cc. 

1 Flask 

1 Funnel, \\ inch 

1 Funnel, 3 inch 

1 Watch glass 

t Evaporating dish, 8 oz. 

t Urinometer 

t Glass rod 

1 Thermometer 

l Crucible, 8 cc. 



1 Piece wire gauze 

l Piece absorbent cotton 

1 Box matches 

l Test tube brush 

1 Test tube rack 

1 Test tube holder, wire 

t Tripod 

1 Piece muslin 

1 Pack filter papers, 3 inch 

1 Pack tilter papers, 6 inch 

1 Sponge 

1 Clay triangle 

2 Tin cans 

1 Copper wash bath 
1 Towel 



Special apparatus, not found in the locker, may be obtained, when needed, 
by handing an order for it to one of the assistants. 




Plate i. Schematic Section of the Kidney. 



1. Cut portion of the Kidney. 2. Pyramid. 3. Papilla of the Pyramid. 4. Glom- 
erulus, enclosed in Bowman's Capsule. 5. Bowman's Capsule. 6. Convoluted por 
tion of Renal Tubule. 7. Loop of Henle. S. Collecting Tubule. 9. Capillary net 
work, — second set ; the first set is the Glomerulus. 10. Openings of Collecting 
Tubules on Papillae, from which the Urine drops into the Pelvis of the Kidney. 



THE SECRETION OF URINE. 



The blood supply to the kidneys is an important consider- 
ation in the secretion of urine. At the outset, one is impressed 
by the fact that a relatively large artery supplies a relatively 
small gland. The blood pressure of the renal artery is nearly, 
if not quite, as great as that of the abdominal aorta, which 
means that about as much pressure is required to force the 
blood through the kidneys as is required to send the blood 
through the pedal extremities. The renal vein is also a rela- 
tively large vessel as compared with the size of the kidney or 
with the efferent vessels of other glands. The pressure within 
the renal vein is very low, practically as low as that in the 
posterior vena cava with which it connects. 

The artery, after entering the kidney, breaks up into 
branches which pass between the pyramids. At the junction 
of the cortex and medulla these branches form arches in the 
substance of the kidneys and from these arches branches run 
outward into the cortex to supply the glomeruli; other branches 
pass inward to supply the pyramids. Each glomerulus has 
its afferent and efferent vessel and of these the efferent is 
smaller. 

On issuing from the capillaries of the glomerulus, the 
efferent vessel soon breaks up into a second set of capillaries 
which supplies the uriniferous tubules ; so that in this arrange- 
ment the blood goes first to the glomeruli and later supplies 
the tubules. The blood from the second set of capillaries is 
finally gathered up by small vessels which unite to form ulti- 
mately the renal vein. The glomeruli and the uriniferous 
tubules are the portions of the kidney actively concerned in 
the production of its secretion — the urine. 

The following points are therefore worthy of special 
notice : in the malpighian body the arterial blood in the glom- 
erular capillaries is separated from the inside of the capsule 
of Bowman by a thin layer of flattened epithelial cells only ; 
two sets of capillaries exist, one set forming the glomeruli and 



the other supplying- the uriniferous tubules, the blood supply- 
ing- the tubules must have first passed through the glomeruli, 
and is therefore more concentrated ; in certain portions of the 
tubules short cylindrical epithelial cells are found which are 
comparable to true secreting- cells found in other glands ; and 
finally, the smaller size of the efferent as compared with the 
afferent vessel fulfills a condition which retards the flow of 
blood through the glomerulus. 

The conditions are favorable for a high and variable 
pressure in the glomerulus and for a lower and more constant 
pressure in the second set of capillaries around the uriniferous 
tubules. On account of the resistance offered by a double 
system of capillaries the blood pressure in the kidneys is kept 
relatively high. 



C < a c t 6 o 




Fig. i. 

l. Artery. 2. Glomerulus. 3. Capsule of Bowman. 4. Convoluted por- 
tion of Tubule. 5. Capillary net work. 7. Loop of Henle. 8. Collecting 
Tubule. 9. Opening of Tubule on Papilla. 



The changes in blood pressure may be observed upon the 
kidney itself by means of an apparatus known as the onco- 
meter. With each rise in the blood pressure the kidney swells, 
with each fall in pressure it contracts, and a tracing can be 
obtained very similar to an ordinary blood pressure tracing. 
The glomerulus suspended in its capsule is also influenced by 
these changes. When the capillaries are engorged with blood 
the glomerulus fills the capsule, when collapsed there is an 
evident space between the membranes of the glomerulus and 
the capsule. 

In 1842 Bowman advanced a theory relating to the pro- 
duction of urine which, while recognizing to some extent cer- 
tain physical factors, also brought out the view that the urine 
is a secretion. 

In 1844 Ludwig advanced the theory, sometimes referred 
to as the mechanical theory, in which only physical processes 
were involved, i.e., filtration, diffusion and osmosis. It is true 
that in the capillaries of the glomerulus there is a high resist- 
ance because of the smaller size of the efferent vessel, there is, 
therefore, a higher degree of pressure in the glomerulus than 
in the capsule in which it is suspended. This inequality of 
pressure is favorable to filtration. As the result of any filtra- 
tion from the glomerulus some of the water of the blood with 
a certain proportion of dissolved salts would pass into the 
beginning of the uriniferous tubule. The effect of such a 
filtration would render the blood remaining in the glomerulus 
and second set of capillaries more concentrated, and in the 
second set of capillaries in connection with the uriniferous 
tubules the essential elements of an osmometer would be 
obtained, — an animal membrane formed by the delicate wall 
of the capillary and the wall of the tubule, upon one side of 
which there is a dense fluid, the blood, and upon the other a 
weak saline solution, conditions which are favorable to the 
processes of osmosis and diffusion. 

In this theory, as a result of the interchange of these 
fluids, some of the water passes from the tubule to the blood, 
making it less concentrated, and the products of retrogressive 
metamorphosis, — urea aud other organic substances, — pass 
from the blood to the tubule, forming urine. An objection in 
connection with the diffusion of urea has been raised in that 




1. Artery. 
Glomerulus. 



Fig. 2. 
2. Afferent Vessel. 



it is a well-known fact that the urine contains a much greater 
amount of urea than does the blood and that it would be con- 
trary to the laws of osmosis and diffusion for a fluid weak in 
urea (the blood) to pass through a membrane a substance 
which has accumulated in greater amount in a fluid (the 

urine) on the other side of 
that membrane. It should also 
be remembered that the blood 
is constantly moving and pre- 
sumably the fluid in the tu- 
bules is doing the same. In 
other words, it would seem 
that if the conditions indicate 
an osmometer, it varies from 
experimental ones in that the 
fluids on either side of the 
membrane, are moving in a 
definite direction. The pro- 
teid constituents do not nor- 
mally leave the blood on ac- 
count of their well-known in- 
disposition to osmosis. 
One important fact, however, remains unaccounted for in 
Ludwig's mechanical theory, and that is, if the uriniferous 
tubules are stripped of their epithelial cells, as they often are 
in disease, urea and some other nitrogenous products are no 
longer, or only imperfectly eliminated, and become stored up 
in the blood and produce the condition known as uremia, 
although the conditions of an osmometer remain. It must, 
therefore.be admitted that there is some direct or elaborating 
action on the part of the epithelium as originally suggested 
by Bowman, although under normal conditions, transudation, 
diffusion and osmotic processes may occur coincidently. 

The theoretical conclusions of Bowman have been con- 
firmed and extended by the practical researches of Heidenhain, 
who injected indigo-carmine into the blood of animals and 
found that it was promptly removed by the kidneys. These 
organs were removed at suitable intervals after the injection 
and carefully examined under the microscope. In no instance 
did he find any of the indigo in the capsules of Bowman, but 



4. Capsule of Bowman. 
5. Efferent Vessel. 6. Capillary net- 
work. 7. Uriniferous Tubule. 8. Vein. 



9 

it was found abundantly in the cells lining certain portions of 
the tubules, and in the lumen of the tubules near these cells. 
Similar experiments with the urate of sodium showed that it 
likewise was secreted at the same place and in the same 
manner. 

Further investigations have tended to diminish the impor- 
tance of the "mechanical" factor and to develop a selective 
or secretory function for the cells, even including- those of the 
glomeruli, by virtue of which the cells will permit certain 
substances to pass through and prevent others, among the 
latter, albumin. The view is advanced that the passage of 
the fluid through the glomerulus is not a mere transudation, 
but a matter of selection also. 

The selective action of the renal cells is both qualitative 
and quantitative. As an example of the former, it is shown 
by experiment that if some egg albumin be injected into the 
blood, it is promptly eliminated by the kidneys. Egg albumin 
is not very markedly different from the serum albumin of the 
blood ; both are indiffusible, but the renal cells recognize the 
former as a foreign substance and immediately separate it 
from the blood. The sugar, in normal quantity in the blood, 
although a diffusible substance is not selected. Urea, which 
is also diffusible, but existing in the blood in much smaller 
amount than sugar, is selected and appears in the urine. Why 
this should be so it is difficult to explain, although it is a well- 
known fact that the sugar serves as a food for the tissues and 
is needed by the system, while urea is a waste product and 
would be detrimental to the system if not eliminated. 

That the cells exert a quantitative selection is shown by 
the fact that when sugar is present in the blood in excessive 
amount, 3 parts per 1000 or over, the excess is promptly 
eliminated. 

Diet influences the reaction of the urine. A vegetable 
diet favors an alkaline reaction ; a diet of flesh favors an acid 
reaction. The urine of the herbivora is therefore alkaline, 
* while in the carnivora and omnivora the urine is mainly acid, 
although influenced to some extent by the kind of food eaten. 
The formation of an acid fluid from alkaline material, — blood 
and lymph, — is at first sight puzzling. The well-known fact 
that the gastric secretion of all mammals is acid is a case in 



10 

point. Furthermore, experimental evidence shows that if an 
alkaline solution of sodium bicarbonate be placed upon one 
side of the membrane in an osmometer and a solution of neu- 
tral sodium phosphate be placed upon the other side and a 
weak electric current be sent through the solutions, the fluid 
in contact with the positive pole will, in a short time, become 
acid from the formation of acid sodium phosphate, while the 
fluid in contact with the negative pole is increased in alkalinity 
Na HC0 3 + Na 2 HP0 4 =Na 2 C0 3 -j-Na H 2 P0 4 . 

While it may be true to a considerable extent that the 
kidney merely removes certain constituents from the blood 
and transfers them to the urine, it has been shown that the 
activity of the renal cells is required in the production of hip- 
puric acid, — a new product, as hippuric acid is not present in 
the blood. 

In general, the theory explaining the secretion of urine, 
according to observed facts, is one which, while recognizing 
the process as partly physical, also requires some process of 
activity or elaboration on the part of the kidney itself. 



II. 

The urine of all mammals may be regarded, for the most 
part, as a solution of constituents derived from the metabolism 
of the tissues of the body. Some of these constituents, espec- 
ially the inorganic, may appear in the urine in the same form 
as they are taken into the body in the food, e.g., sodium 
chloride ; others, especially the organic, represent decom- 
position products derived from the food or tissues, e. g. y urea, 
creatinin, etc. The composition of the urine may, therefore, 
to some extent be regarded as an index of tissue activity, and 
the examination of this secretion is of considerable importance 
in clinical diagnosis and prognosis. 

That there is a relationship between the diet and the 
renal excretion is shown by the examination of the urine of 
the three great classes of animals grouped according to the 
food they eat ; herbivora, omnivora and carnivora. Perhaps 
the first and most striking characteristic of the urine of each 
group is its chemical reaction. In vegetable feeders it is nor- 



11 

mally alkaline ; in flesh eaters it is markedly acid ; and in the 
omnivora it may be acid or alkaline according to the prepon- 
derance of the fleshy or vegetable material in the food. 

In the practical treatment of this subject it is convenient 
to regard the horse as the type for the herbivora ; man for 
the omnivora ; and the dog or the cat for the carnivora. In 
all cases throughout this work it is to be understood that, 
unless especially directed, the same tests are to be performed 
upon the urines of man and horse and a parallel record of such 
tests, for comparison, is to be kept upon the blank pages. 

In man, fresh, normal urine is a clear, amber-colored, 
transparent liquid, having a peculiar aromatic characteristic 
odor, and a bitter saline taste. 

In the horse, the urine is of a yellowish color when passed, 
but turns to a deep brown color upon standing for a time, due 
to the oxidation of pyrocatechin. It is more or less turbid and 
of a viscous character. Its odor is somewhat ammoniacal and 
strongly aromatic and more penetrating than that of man. 

The urine is chiefly a solution of urea and certain organic 
and inorganic salts ; epithelial cells and mucus may also be 
held in suspension. Like milk and other animal fluids, the 
urine is not of constant composition. It is influenced by vari- 
ous factors, such as food, the amount of water or other fluids 
taken into the body ; the temperature of the skin ; the emo- 
tions ; blood pressure, general or local ; exercise ; the time of 
day ; age ; sex ; and medicines. 

Quantity. The amount varies considerably. In man, 
the quantity for twenty-four hours ranges from 1000 cc. to 
2000 cc. In the horse, the average amount is about 3000 cc. 
to 4000 cc, although it may go as high as 7000 cc. or 9000 cc. 
In the ox a still greater quantity is secreted, the usual limits 
being from 4500 cc. to 19000 cc. In the sheep, from 250 cc. to 
700 cc. In the pig it varies from 1200 cc. to 6000 cc. In the 
dog it varies from 200 cc. to 900 cc, depending upon the size 
of the animal. 

Color. The color ranges from pale yellow to brown. 
The normal color of urine is due to pigments probably derived 
from the coloring matter of the bile. 

Transparency. The urine of the horse is normally more 
or less opaque, that of man should be transparent at the time 



12 

of passing. Many pathological urines, however, are perfectly 
clear. Patholog-ical turbidity may be due to urates, phos- 
phates, pus, bacteria, spermatozoa, fatty globules, blood, etc. 
All urines become turbid after standing- for a time. 

The urine of the horse is especially turbid at the end of 
micturition, exceptionally it may be clear but becomes turbid 
on cooling-. Its opacity is due to the presence of earthy salts 
of the carbonate of lime precipitated and formed by the dis- 
eng-ag-ement of a certain amount of carbon dioxide from the 
bicarbonate of lime. The turbidity increases when the urine 
remains for any leng-th of time in the bladder ; it reaches its 
maximum when the urine is cooled by exposure to the free air; 
it diminishes after the iog-estion of a larg-e quantity of water. 

A clear, limpid urine is g-enerally patholog-ical in the 
horse ; it indicates polyuria and the reaction in this case is 
usually acid, exceptionally neutral or alkaline when the phos- 
phates are modified qualitatively or quantitatively. 

The turbidity of horse urine is abnormal when it is due 
to the presence of the phoshate of lime, or of calcium sul- 
phate, or of urea and acid salts ; also from the existence of 
albuminoid substances, (exudates, leucocytes in interstitial 
nephritis). 

In other animals the passing- of turbid urine is usually 
reg-arded as abnormal. 

Consistency. In the horse the urine is very viscid on 
account of the contained mucus and sometimes of epithelial 
debris. It filters very slowly. Urine taken directly from the 
ureter or the pelvis of the kidney is still more viscid, having- 
a consistency very similar to that of eg-g- albumin and a spe- 
cific gravity somwhat higher than normal. 

Reaction. The reaction of human urine is acid, that of 
the dog- more so than that of man. In the pig- it is sometimes 
acid and sometimes alkaline, depending- upon the diet. In the 
horse and sheep it is alkaline, also in the ox, but in the calf 
and foal it is acid. Herbivorous urine is alkaline, but if such 
animals are starved for a time they practically become car- 
nivorous in that they are living- upon their own tissues and 
under such conditions their urine becomes acid. 

In the herbivora the reaction is normally alkaline and is 
due to the presence of bicarbonates or carbonates of lime or 



13 

potassium. It would appear that the salts present in the 
vegetable food undergo oxidation to form organic acids, and 
these in turn are transformed into bicarbcnates or carbonates, 
causing alkalinity of the urine. 

In man the acidity is due to the presence of acid sodium 
phosphate NaH 2 P0 4 . The urine passed before breakfast and 
during fasting or perspiration is more acid than at other times ; 
during digestion and after meals the acidity is decreased. 

In man there is increased acidity physiologically during the 
night ; with a flesh diet ; after strong muscular exertion ; dur- 
ing the intervals of gastric digestion ; after the ingestion of 
mineral acids. 

There is increased acidity, pathologically, in fevers ; in rheu- 
matism ; after asthmatic attacks ; in emphysema, pneumonia 
and pleuritis. 

The urine is less acid, or alkaline, physiologically, during 
gastric digestion ; after hot or prolonged cold baths ; after 
profuse sweating ; after copious ingestion of vegetable acids 
and their salts. 

Pathologically in acute and chronic inflammation of the urin- 
ary tract as in cystitis ; in decomposition of the urine in the 
bladder in retention ; in some cerebral and nervous diseases ; 
in anemia ; chlorosis ; and general debility. 

When the urine of man is set aside in a cool place it grad- 
ually becomes more acid. This is called acid fermentation. 
After longer exposure to a warm atmosphere the urine becomes 
neutral, and finally strongly alkaline in reaction. It becomes 
turbid, has an ammoniacal odor, and deposits triple phosphate, 
ammonium urate, and great numbers of microbes exist. This is 
alkaline fermentation and is due to the transformation of the 
urea into ammonia and carbon dioxide, by means of a ferment 
produced by an organism known as the micrococcus ureae. (Fig. 
20). This organism is said to be conveyed through the air and 
to exist commonly around the orifice of the urethra. As long 
as the urine is acid the organism does not exist in the bladder, 
but may sometimes gain entrance through the medium of a 
sound or catheter. 

Test the reaction of the urines with red and blue 
litmus paper. Some urines change both the red and 
blue paper and are termed amphoteric. (In taking notes 



14 



of the experiments it is well to record the results in 
parallel columns, the horse urine in one column and the 
human urine in the other). 
Specific Gravity. The specific gravity of human urine 
ranges from 1015 to 1025, the average being 1018 to 1020. 
That of the horse ranges from 1020 to 1050, the average being 
about 1035. That of the ox is lower, ranging from 1015 to 
1045. It seems to depend considerably upon the milk secreted. 
In milch cows the urine contains a greater amount of water 
and a lesser amount of solids. The specific gravity of the 
urine of the sheep ranges from 1015 to 1060 ; that of the pig 
from 1005 to 1025. and of the dog from 1016 to 1060, depend- 
ing upon the diet. Cat, 1020 to 1040. 

The specific gravity may be ob- 
tained in different ways. The sim- 
plest and most usual way is to employ 
the instrument known as the urin- 
ometer. Some urinometers are not 
strictly accurate, but they may be 
tested by filling the urinometer jar 
with distilled water at 15° C. (60° F.) 
Read the division of the scale corre- 
sponding with the surface of the 
fluid looking above or below the 
meniscus as is found to be the most 
correct for the zero reading. Always 
adhere to this method when using 
the same urinometer. Test the spe- 
cific gravity of the urines and make 
the necessary corrections. If the 
urine is warmer than 15° C. add 1 to 
the last right hand figure of the spe- 
cific gravity for every 4 degrees of C. 
temperature, or for every 7 degrees 
of extra F. temperature. As oppor- 
tunity presents, test the specific grav- 
ity of some warm, freshly passed 
urine. Test the same urine later, 
when cool, and note if any difference 
in the reading. 




Fig. 3. 
Urinometer. 



15 

Urinometers already corrected for the ordinary room tem- 
perature (70° F.) may be obtained, in which case the temper- 
ature corrections may be omitted. If the urine should be too 
dense to read easily on the urinometer, dilute it with an equal 
volume of distilled water and multiply the reading by two to 
get the correct specific gravity. The variation of the specific 
gravity depends upon the amount of the solids in the urine. 
The amount of solids may be estimated with approximate 
accuracy from the specific gravity by Christison's formula 
(Haser-Trapp's coefficient): " Multiply the last two figures of 
a specific gravity expressed in four figures by 2.33. This 
gives the quantity of solid matter in every 1000 parts." (The 
number of grams in 1000 cc). 

Example. Suppose a patient passes 1400 cc. of urine in 
24 hours and the specific gravity is 1020, 20X2.33=46.6 grams 
of solids in 1000 cc. To ascertain the amount in 1400 cc. use 
the following proportion: 1000 cc. : 1400 cc. :: 46.6 grams is 
to X (65.24 grams). The total quantity of the solids of the 
human urine is about 60 grams for the 24 hours or approxi- 
mately 4%. 

The following method taken from the Alkaloidal Clinic is 
also given : Multiply the quantity, in ounces, of the 24-hour 
urine by the last two figures of the specific gravity and this 
by 1.1, the product will represent the total solids in grains. 
Thus, if the amount of urine voided in 24 hours be 36 ounces 
and its specific gravity 1021, the formula would be 36X21X1.1, 
equal to 831 grains, the normal amount for a person weigh- 
ing 100 lbs. The amount for other weights may be determined 
by proportion. 

In general the amount of total solids is a measure (a) of 
the activity of tissue change ; (b) of renal integrity ; (c) of 
abnormal constituents in the urine. Hygienic conditions 
which favor increased metabolism, as abundant food, active 
exercise, etc., increase the solid matter in urine, while the 
opposite conditions decrease them. 

With the urine normal or subnormal in amount, the solids 
are deficient pathologically from defective and enfeebled metabolism 
as in senility ; anemia as a result of syphilis, cancer, etc. ; 
chronic alcoholism ; functional or organic diseases of the liver. 
From renal failure as in acute nephritis ; certain conditions 



16 

of chronic renal disease ; at the close of Bright's disease ; 
venous congestion of the kidneys, etc. With the urine in- 
creased in amount there may be a deficiency of solids in dia- 
betes insipidus ; interstitial nephritis ; amyloid disease of the 
kidney ; chronic parenchymatous nephritis. When the urine 
is not increased in amount, the urinary solids are increased in 
fevers ; lithemia ; some forms of dyspepsia. When the quan- 
tity of urine is increased the solids are increased in diabetes 
mellitus; phosphaturia ; azoturia (excessive secretion of urea). 
Calculate the solids of the urines by the formulae 
previously given. After obtaining the result for 1000 cc. 
estimate the quantity in 1250 cc. of human, and 5450 cc. 
of horse urine. In recording the tests use parallel col- 
umns, one column for the horse and the other for the 
human urine. 



III. 



Qualitative Tests. In all cases the urines must be filtered 
and perfectly clear before attempting the examination. 



INORGANIC CONSTITUENTS. 

These consist chiefly of sodium, potassium, ammonium, 
calcium, and magnesium, combined with hydrochloric, phos- 
phoric and sulphuric acids. 

Water. The water of the urine is derived from the food 
and drink, a small quantity being formed in the body. It var- 
ies according to the activity of the sweat glands of the skin. 

Chlorides. Next to the urea the chlorides form the chief 
portion of the urinary solids. The chlorides are increased 
physiologically after the ingestion of salt foods and much 
water ; mental and physical activities ; and during pregnancy. 
Pathologically, they may increase after the crises of fevers ; 
after the absorption of exudates; in diabetes (occasionally). 

The chlorides are decreased pathologically, in all acute 
fevers ; pneumonia (often entirely absent during the height 
of the disease); in cholera ; and in most chronic diseases. An 
increase, or the re-establishment of the excretion of chlorides 



17 

in disease is generally a favorable sign. In pneumonia it is 
a precursor of the crisis, and may often take place before other 
symptoms reveal the favorable change. 

Test a portion of each urine with a few drops of 
silver nitrate solution. A white, cheesy or curdy pre- 
cipitate insoluble in nitric acid indicates the presence 
of silver chloride. The phosphate of silver may also 
be thrown down but the nitric acid dissolves it, keeping 
it in solution. 

Evaporate carefully a few drops of urine upon a 
glass slide, with a gentle heat over the flame. Octa- 
hedral or rhombic crystals may form, — a compound of 
sodium chloride and urea. Examine with the micro- 
scope. (Fig. 5). 
Sulphates. The sulphates are chiefly those of sodium 
and potassium. Only a small amount of them enters the body 
with the food, so that they are chiefly formed from the meta- 
bolism of proteids in the body. The above are known as 
ordinary sulphates. Another class known as the ethereal sul- 
phates also exist. The proportion exists in the ratio of 10 of 
the ordinary to 1 of the ethereal in man. In the horse the 
proportion is about 2 of the ethereal to 1 of the ordinary. 
The ethereal sulphates are formed by the combination of sul- 
phuric acid with organic bases such as phenol, skatol, etc., 
which originate from putrefactive processes in the intestine. 
The amount of ethereal sulphates is of importance in deter- 
mining whether or not the digestive processes are going on 
normally. In general the sulphates are increased physiolog- 
ically by the ingestion of sulphur and its compounds ; nitro- 
genous food ; and conditions of increased metabolism. 

After acidulating the urine with hydrochloric acid 
to prevent the precipitation of phosphates, add to a small 
part of each urine, a little 2% barium chloride solution ; 
a precipitate of barium sulphate is formed, insoluble in 
nitric acid. 

To separate the ethereal sulphates, mix 30 cc. of 
urine with an equal bulk of " baryta " mixture. Stir and 
filter. This removes the ordinary sulphates (as barium 
sulphate), add 10 cc. of hydrochloric acid to the above, 
filtrate, and keep in the water bath at 100° C. for an 



18 

hour in the hood and then allow the ethereal sulphates 
to settle. This may require some little time. (Baryta 
mixture is prepared by making- saturated solutions in 
the cold, of barium nitrate and barium hydrate, and 
adding- two volumes of the hydrate to one volume of the 
nitrate). 
Phosphates. The phosphates consist of alkaline and 
earthy salts in the proportion of 2 to 1. The latter are insol- 
uble in an alkaline medium and are precipitated when acid 
urine becomes alkaline. They are insoluble in water, but 
soluble in acids ; in acid urine they are held in solution by free 
C0 2 . The alkaline phosphates (sodium and potassium) are 
very soluble in water, and they never form urinary deposits. 
The eaithy are phosphates of calcium (Ca 3 P0 4 ) 2 (abundant) 
and magnesium (MgHP0 4 plus 7H 2 0) (scanty). An alkaline 
medium precipitates them although not in the form in which 
they occur in the urine. 

The excretion of phosphates in the urine is larg-ely de- 
pendent upon the amount of calcium ing-ested ; the more cal- 
cium the food contains, the less phosphoric acid appears in the 
urine, and the more in the feces. This is due on the one hand 
to the tendency on the part of calcium to form insoluble cal- 
cium phosphates in the intestinal tract and in this way to 
prevent the absorption of the food phosphates ; on the other 
hand, to the well-established tendency of calcium salts to be 
excreted into the bowel and not into the bladder ; one must 
imagine in the latter case that calcium salts circulating- in the 
blood combine with circulating phosphoric acid and bear the 
latter with them into the bowel. 

The fact is of some therapeutic importance in the treat- 
ment of nephrolithiasis due to uric acid calculi, for the admin- 
istration of calcium salts in this affection by bearing - much 
phosphoric acid into the bowel, leads to the excretion of less 
phosphoric acid in the urine, and hence of normal and basic 
instead of acid phosphates ; and as the latter precipitate and 
the former dissolve uric acid, it will be seen that by giving 
calcium we prevent the precipitation of crystalline uric acid 
and urate deposits in the urinary passag-es. (Croftan). 

Where considerable calcium is present in the food the 
excretion of phosphates in the urine is minimum, especially if 



19 

the urine is alkaline because of the presence of sodium or 
potassium salts. In herbivorous animals where the urine is 
alkaline and where considerable quantities of phosphorus con- 
taining food are eaten, very little phosphate is excreted in 
the urine. 

Phosphates are increased in the urine pathologically in 
rickets; osteomalacia; osteoporosis; fractures; chronic rheu- 
matism ; diseases of the nervous system ; and after great 
mental strain and worry. Phosphates are decreased in renal 
diseases and phthisis. 

Physiologically, variations occur chiefly from the character 
of the food and drink ; in the horse there may be an increase 
in the urinary phosphates after a large feed of oats, bran, oil- 
cake, etc. Pathologically, they are increased during the active 
changes in such bone diseases as spavin, ring-bone, splint, etc. 
To a small amount of each urine add about half its 
volume of nitric acid and then add two volumes of a 5% 
solution of ammonium molybdate and boil A canary 
yellow ppt. (crystalline) of ammonium phospho-molyb- 
date should appear in the omnivorous urine. In the 
herbivorous urine the presence of so much organic mat- 
ter renders the test unreliable ; although a precipitate 
may form it is not a typical or characteristic one for 
phosphates in the urine of the horse, unless the organic 
matter has been previously removed. 

To each of the urines add half its volume of am- 
monia and allow it to stand. A precipitate of earthy 
phosphates is formed, in the urine of man. Filter, add 
enough nitric acid to give an acid reaction to the urines, 
and test the filtrates with ammonium molybdate as 
before. This method separates the earthy from the 
alkaline phosphates. 

To a portion of the urines add half their volumes 
of baryta mixture ; a copious precipitate. Filter, add 
nitric acid and test the filtrates with ammonium molyb- 
date. No ppt. should occur as the baryta mixture pre- 
cipitates the phosphates as well as the sulphates and 
carbonates. 

Use a little of the magnesia mixture instead of the 
baryta mixture. Filter, add a little nitric acid, and test 



20 

the filtrate with ammonium molybdate. (The magnesia 
mixture is composed of magnesium sulphate 1 part, 
ammonium chloride 1 part, ammonia water 1 part, and 
distilled water 8 parts). 

To portions of the urines add a few drops of acetic 
acid and then a little $'/<> uranium nitrate solution, — a 
yellow ppt. of uranium phosphate is formed. 
The lime, magnesia, iron and other inorganic urinary 
constituents are comparatively unimportant, and have no 
special clinical significance. The tests for them are some- 
what complicated and are therefore omitted. 

Demonstration of Carbonates and C0 2 in Urine. CO z 
exists in the urine to some extent in a free state. There are 
also various carbonates present, especially in the herbivorous 
urine. These may be broken up and C0 2 given off by the 
application of heat or certain acids. 

Heat experiment. Fill a small flask about half full 
of unfiltered herbivorous urine. Through the perfor- 
ated stopper of the flask pass some bent glass tubing 
connected with a test tube containing lime water. Heat 
the urine in the flask, and as it boils the CO a will pass 
over into the test tube, and calcium carbonate will be 
formed from the union of the gas with the lime. Repeat 
the experiment with omnivorous urine. 

A simple proof of the presence of carbonates, or 
C0 2 in the urine is to fill a Doremus Ureometer with 
25 cc. of the unfiltered urine and introduce 1 cc. of nitric 
acid. The larger part of the gas (C0 2 ) rises and col- 
lects in the upper portion of the ureometer. Compare 
the amounts thus obtained in the urines of horse and 
man. 

A simpler qualitative test is to add a few drops of 
nitric or acetic acid to a little unfiltered urine in a test 
tube. If effervescence occurs it is due to C0 2 set free 
from the carbonates by the acid. 



21 



IV. 



ORGANIC CONSTITUENTS. 

Urea is, in amount, the principal constituent of the solids 
of the urine. It is the most important product of the decom- 
position of proteid in the food. In round numbers it forms 
from 2% to 3.5% of the urine, averaging- about 2.5%. About 
one-half of the total solids in the urine consists of urea. 
Urea has no effect on litmus, it is odorless, has a weakly cool 
and bitter taste like saltpeter. It is very soluble in water and 
alcohol, but it is almost insoluble in ether and benzine. About 
90% of the total nitrogen of the urine is excreted in the form 
of urea. 

Physiologically, urea is increased by a proteid diet ; exercise 
and muscular vigor ; by drinking much water. It is decreased 
by fasting ; non-nitrogenous food ; reduction of water in the 
diet ; alcoholic beverages, tea or coffee ; indolence of mind and 
body. Pathologically, it is increased in all acute fevers ; dys- 
pnoea ; diabetes ; and phosphorus poisoning. It is decreased 
in uremia ; acute yellow atrophy of the liver and in chronic 
diseases. In general, any disease that interferes with the ac- 
tivity of the liver decreases the urea. Any disease affecting 
the uriniferous tubules may modify the appropriation of the 
urea from the blood and affect its passage into the urine. An 
increase of the urea independently of the physiologic variations 
and amount of nitrogenous food eaten is an approximate index 
of the amount of tissue waste in the system ; on the other 
hand, when the urea is decreased it is an evidence of a diseased 
condition of the liver (the producer of urea) or the kidney (the 
eliminator of urea). 

A simple method of detecting urea is to concen- 
trate a small amount of human urine in an evaporating 
dish to about half of its original volume. Place a drop 
or two of this concentrated urine upon a glass slide and 
after adding a drop of nitric acid, gently warm over the 
flame. If urea be present, upon evaporation, the micro- 



22 




Fig. 4. 
Crystals of Nitrate of Urea. 



scope will show the charac- 
teristic crystal of nitrate of 
urea, of rhombic or hexag- 
onal form. (Fig - . 4). 

Take 20 cc. of fresh, fil- 
tered omnivorous urine and 
add 20 cc. of baryta mixture 
to precipitate the phosphates 
and sulphates. Filter, evap- 
orate the filtrate to dryness, 
and extract the residue with 
a little boiling- alcohol over 
the water bath very care- 
fully. Filter off the alcoholic 
solution, place some of it on a 
slide and allow the crystals of 
urea, usually long, fine, trans- 
parent needles, to separate out. 
Examine them under the micro- 
scope. ( Fig. 5). 

Repeat in the hood, the experi- 
ment upon the urine of the horse 
and compare results. 

Heat some urea crystals in a 
test tube. Biuret is formed and 
ammonia comes off. Add a trace 
of copper sulphate solution and a 
few drops of 20% caustic potash. A rose-red color is 
produced, — the biuret reaction. 
Medicines which increase the amount of urea are : urea 
itself ; uric acid, common salt, phosphoric acid, squill, theo- 
bromine, colchicum, cubebs, atropine, cantharides, vegetable 
acids, iron preparations, Hyposulphite of soda, potassium chlor- 
ide, ammonium chloride, coca, potassium permanganate, oxy- 
gen, salicylic acid. 

Medicines which decrease the amount of urea are : digi- 
talis, alcohol, coffee, tea, potassium and sodium iodides, potas- 
sium bromide, arsenic, turpentine, alkaline carbonates, mercury, 
antipyrin, valerian, quinine sulphate, benzoic acid. 




Fig. 5. 
Crystals of Urea. 



23 

Uric Acid, sometimes called lithic acid, is, next to urea, 
the most important nitrogenous constituent of the urine of 
man. It has been said to be absent in herbivorous urine, being- 
replaced by hippuric. This has been shown by later researches 
not to be strictly true, as a trace of uric acid is found in 
addition to the hippuric. In birds and reptiles uric acid is the 
chief nitrogenous constituent, being present in greater amount 
than urea. 

In man the proportion of urea to uric acid is about 45 to 
1, about 0.5 gram of the latter being excreted in the 24 hours. 
It causes the brick red deposit sometimes seen in urine after 
standing for a time. — Its solubility is very low, only 1 part 
being soluble in 14000 cc. of cold water, and 1 to 18000 in 
boiling. Physiologically, it is increased and diminished propor- 
tionately with urea. Pathologically, it is increased in indigestion ; 
acute dropsies, rheumatic and catarrhal inflammations ; after 
attacks of gout ; cancer of the liver ; in leucemia ; in all dis- 
turbances of the circulation and respiration. Pathologically 
uric acid is decreased in chronic diseases generally ; diabetes 
and polyuria ; before paroxysms of gout ; anemias ; chronic 
rheumatism ; chronic diseases of spinal cord. 

Uric acid is generally in solution in the form of urates of 
sodium, potassium, ammonium, lime and magnesium. These 
salts are very easily decomposed even by weak organic acids. 
Perform the following tests upon both urines (filtered). 
In a conical glass, add 5 parts of hydrochloric acid 
to 30 parts of urine. Label and put in a cool place for 
24 hours. Red or brownish colored crystals of uric acid 
are deposited upon the sides of the glass, or form a 
pellicle on the surface of the fluid like fine grains of 
cayenne pepper. The brownish-red color is due to pig- 
ment (uroerythrin). 

Murexide Test. To about 1 cc. of human urine add 
a little nitric acid ; evaporate in a porcelain dish very 
carefully to avoid charring. Cool and add a drop of 
ammonia, a purple red color of murexide or purpurate 
of ammonia is formed. It turns bluer upon the addition 
of caustic potash solution. 

Dissolve a few crystals of uric acid in 10% caustic 
soda or potash. Add a drop or two of Fehling's Solu- 



24 



tion — or dilute cupric sulphate and caustic potash and 
heat — there should occur a ppt. which at first may be 
white and after a time turning- green or reddish. 

(Fehling's Solution is put up in two bottles; one 
labeled A, the other B. In making tests, take equal 
parts of A and B and add the substance to be tested). 

Schiff's Test. Dissolve a little uric acid in a small 
quantity of 10% sodium carbonate solution. With a 
glass rod, place a drop of silver nitrate solution on filter 
paper and then a drop of the uric acid solution so that 
the two drops partially overlap. A dark brown or black 
spot of reducing silver appears. 
Hippuric Acid, (C 9 H 9 N0 3 ), is found especially in the 
urine of the herbivora, as the horse, ox, etc. In the urine of 

carnivora, and especially in that 
of man, it exists in but very min- 
ute quantity, usually about 0.5 to 
1 gram being excreted in 24 hours. 
It dissolves readily in hot alcohol 
but is sparingly soluble in water. 
It occurs in man after the inges- 
tion of certain vegetables, such as 
asparagus, plums, pears, and ap- 
ples with their skins ; a purely 
vegetable diet and from the use 
of benzoic acid, cinnamic acid, 
essence of bitter almonds, quinine, 
and analogous bodies. Hippuric 
acid normally seems to be derived 
chiefly from the husks or cuticular 
structures of the food. 

Pathologically hippuric acid is in- 
creased in diabetes, chorea ; jaun- 
dice and other liver complaints ; 
and in the acid urine of patients 
suffering from all kinds of fevers. 
In testing for hippuric acid the 
fresh urine should be used ; if 
Various forms of Hippuric Acid stale, benzoic acid is likely to be 
with triple phosphates. obtained instead. 




Fig. 6. 
Crystals of Hippuric Acid. 




FiG 



25 

Test. Saturate the fresh urine with lime water, 
which transforms the hippuric acid into a salt of lime, 
the fluid is then filtered, evaporated to a syrupy consist- 
ency and excess of hydrochloric acid added, when hip- 
puric acid crystallizes out on standing-. The horse 
urine is to be evaporated in the hood. 
Creatinin (C 4 H 7 N 3 0). This substance was discovered 
in the urine by Liebig. It is easily produced from creatin, a 
substance normally existing- in plain and striated muscular 
tissue. Creatinin occurs constantly in normal human urine, 
the amount varying according to Voit, from 0.5 to 4.9 grams 
per day according to the quantity of proteids eaten. It is 
said not to be diminished by fasting. 

Pathologically it is increased in 
typhoid fever ; intermittent fever; 
pneumonia ; tetanus. It is de- 
creased during convalescence from 
the above diseases ; and likewise 
in anemia ; chlorosis ; muscular 
atrophy ; tuberculosis ; paralysis, 
etc. 

Test. Add a little caustic 
soda solution to the urine and 
F,G - 8 - then a few drops of freshly 

prepared \°fo sodium nitro- 
t prusside. A ruby red color develops. Boil, the color 

fades. While boiling, add a little acetic acid, the color 
changes to blue. The above is a modification of Weyl's 
test. Acetone, if present, also gives the red reaction. 
Perform the above test on both urines. 
Caution Creatinin reduces copper oxide and may 
be taken for small quantities of sugar. 
Mucus. Mucus in the urine is not visible, but causes 
cloudiness sometimes by entangling epithelial cells, urates, 
oxalate of lime and other crystals in various amounts. 

Add to the urine a little acetic or citric acid and, 
in addition, a few drops of liquor iodi comp. (Lugol's 
solution) which makes the threads or bands of mucin 
visible. 




26 

Indican orllndoxyl. This substance is derived from 
iudol, one of the putrefactive products formed in the intestine. 
Indoxyl occurs in very small quantities in normal human urine, 
about .004 to .020 gram for the 24 hours. Horse's urine is 
said to contain 23 times as much. The intestines of the herb- 
ivora are much longer than in the case of the carnivora. On 
this account, and in conjunction with the carbohydrate diet, 
a much greater fermentation occurs, which leads to a greater 
elimination of indican in the urine. 

In obstruction of the intestine, or in intestinal catarrh or 
where the food remains a long time in the intestine and fer- 
ments there, the proportion of indican increases in the urine 
and causes a true indicanuria. Indoxyl is of considerable 
clinical importance, an increase is indicative of imperfect per- 
formance of the digestive processes. In obstructive diseases 
of the small intestine the increase of indoxyl in the urine is 
enormous. 

Pathologically indoxyl is increased in cholera, typhoid fever, 
peritonitis, dysentery, Addison's disease, cancer of the liver 
and stomach and pernicious anemia. 

Jaffe's Test. To a little urine add an equal volume 
of strong hydrochloric acid. Add to this mixture 2 or 
3 drops of a solution of freshly prepared chlorinated 
soda. There soon forms a bluish cloud of indigo. Add 
a little chloroform, this will take the indigo into solu- 
tion and settle as a blue layer at the bottom of the test 
tube. The amount of indoxyl can be judged by the 
depth of the blue color. Indican is oxidized by free 
chlorine obtained from the chlorinated soda to indigo. 
The hydrochloric acid — ferric chloride test. This 
reagent has the advantage of keeping indefinitely. It 
is prepared by dissolving 2 grams of solid ferric chloride 
in 500 cc. of concentrated hydrochloric acid. 

To equal parts of urine and the above reagent add 
a little chloroform. Shake frequently but not too vio- 
lently (otherwise an emulsion may be formed). The 
chloroform will become more or less blue by the indigo 
formed, in proportion to the indican originally present. 
There is no exact quantitative method for indican. 
• According to the depth of the blue color it may be des- 
ignated as little,. much or copious. 



27 

Oxalic Acid. This is usually found in combination with 
lime in the form of calcium oxalate. The crystals are of small 
size and appear in the form of dumb bells and octahedra. 
They occur normally in greater amount in herbivorous than 
in omnivorous urine. They greatly increase after eating such 
vegetables as tomatoes, fresh beans, beet-root, asparagus, 
apples, grapes, honey, and after the use of rhubarb, senna, 
squills, etc. Another source of oxalic acid in the body is 
incomplete oxidation of carbohydrates and proteids or retarded 
metabolism. It is therefore a result of mal-assimilation and 
is found in dyspepsia, diabetes mellitus, etc. The long con- 
tinued excretion of an excess of oxalate of lime frequently 
irritates the kidneys, producing albuminuria and grave nervous 
disturbances and may lead to the formation of calculi. (Fig. 18.) 
Acetone. Normal urine may contain traces of acetone 
but it occurs in excessive quantities as a pathological con- 
dition. It is found in many of the fevers, certain forms of 
cancer, in starvation, and in diabetes, when it indicates an 
advanced form of the disease. It is associated with an in- 
creased proteid metabolism and is looked upon as a product of 
proteid decomposition with deficient oxidation. 

Lieben-Ralfe Test. Dissolve 1.3 grams (20 grains) 
of potassium iodide in 4cc. (1 dram) of liquor potasse. 
Boil in test tube, after which gently pour the urine on 
its surface. A yellow precipitate between the two solu- 
tions indicates an affirmative test. Or to the urine add 
a few crystals of iodine and of iodide of potassium with 
some caustic potash. Heat. Yellow precipitate — iodo- 
form with its characteristic odor. 

Legal's Test for Acetone. Add to 5 cc. of the urine 
some fresh aqueous solution of sodium nitroprusside 
followed by a little ammonia, or sodium hydrate solu- 
tion, which gives a red color disappearing on boiling. 
Add sufficient glacial acetic acid and a purple or violet 
red color results. Compare with creatinin. 

These tests are not always satisfactory when ap- 
plied to the ordinary urine. Greater accuracy is claimed 
if the urine is distilled and the tests applied to the 
distillate. 



28 

Sternberg's Test. Acidulate the suspected fluid 
with a few drops of phosphoric acid, and then add small 
quantities of solution of copper sulphate, and of iodine 
in potassium iodide (Lugol's Solution) ; when acetone 
is present a brown cloudiness appears ; on heating - , the 
liquid is decolorized, and a grayish-white, pulverulent, 
voluminous precipitate appears, which contains iodine 
and copper in organic combination. The precipitate is 
almost insoluble in water. Alcohol affords a similar 
reaction, but only after prolonged boiling, and gives a 
sparing precipitate. 

Frommer's Test for Acetone. To 10 cc. of urine in 
a test tube add 1 gram of solid potassium hydroxide ; 
before the latter is dissolved add 10 to 12 drops of sali- 
cylic aldehyde (made by dissolving 1 part of salicylic 
acid in 10 parts of absolute alcohol). Heat the mixture 
to about 70° C. In the presence of acetone, there is 
formed a scarlet red ring. According to Frommer, even 
the minutest amount of acetone will give this reaction 
and no other constituent of the urine will give this color 
— not even diacetic acid. The reaction is explained as 
follows : One molecule of salicylic aldehyde combines 
with one molecule of acetone to form oxybenzol-acetone. 
This, in the presence of strong alkalies, forms dioxy- 
dibenzol-acetone. The alkaline salts of this compound 
are intensely red. 
Urobilin is commonly regarded as the most important 
coloring matter in the urine. There is some evidence that it 
represents a reduced form of bilirubin, one of the pigments of 
the bile. Urobilin is more readily obtained from highly col- 
ored urines, (fevers, etc. ). 

Test. The ordinary test with an alcoholic solution 
of zinc may be simplified in the following manner : 
10 cc. of urine are acidified with 2 drops of HC1 and 
shaken with 2 cc. of chloroform. After the separation 
of the liquids, 2cc. of the chloroform layer are tested 
with 4cc. of a solution of 1 gram of crystallized acetate 
of zinc in a liter of 95% alcohol (shelf reagent). At 
the junction of the two layers the green fluorescent ring 
characteristic of urobilin will appear, and on shaking, 



29 

a fluorescence which is rose-colored by reflected light 
will be distinguished throughout the liquid. 

Another test is to add ammonia to the urine until 
distinctly alkaline, filter, and to the filtrate add a little 
chloride of zinc solution. A green fluorescence should 
appear, and if examined with the spectroscope, a char- 
acteristic band should occur. (See Fig. 16). 
Leucin and Tyrosin are pathologic constituents of urine. 
They are normal products of pancreatic digestion and under 
ordinary conditions are carried, after absorption, to the liver, 
where they disappear, presently undergoing decomposition. 
When present in the urine these bodies are usually considered 
pathognomonic of acute yellow atrophy of the liver, although 
they are likewise stated to be present in the urine in certain 
rare cases of acute phosphorus poisoning associated with 
hepatic atrophy, due to typhoid fever, etc. 

Phenol. According to Tereg and Munk the horse excretes 
in the urine about 10 grams of tribromphenol in 24 hours. 
The tribromphenol is equivalent to 3 grams of phenol daily. 
Great importance is laid by these observers on the excretion 
of phenol, a process which is suspended during intestinal com- 
plaints, particularly colic, and is, according to them and others, 
a cause of the rapid death in these affections, produced by the 
toxic effect of the unexcreted phenol. The production of 
phenol in the healthy body is greatly influenced by diet, being 
largest on rye and hay, one part peas and two parts oats, and 
on hay alone ; it is smallest on rye alone, and next smallest 
on oats and hay. Salkowski is inclined to regard the excretion 
of 3 grams of phenol daily as too high. 



V. 

ABNORMAL SUBSTANCES FOUND IN THE URINE. 

Albumin. The presence of this substance in the urine is 
regarded as pathologic. There is, however, in the urine of 
some individuals apparently enjoying perfect health, minute 
traces of albumin sometimes present, and, unless these traces 
persist, are not to be regarded as serious. If present in any 
considerable quantity, it must be regarded as distinctly abnor- 



30 

mal. Albuminuria is the term applied when albumin occurs 
•in notable quantity in the urine. The principal form of albu- 
min present is serum-albumin, in addition there may be serum- 
globulin, acid albumin, albumose, and peptone. 

The amount of albumin in the urine may be increased : 
1. By food rich in albumin ; 2. Suppression of cutaneous per- 
spiration, as by colds, burns, or cutaneous diseases ; 3. Pul- 
monary and cardiac diseases attended with dyspnoea, cyanosis, 
valvular disease of the heart ; 4. Febrile and inflammatory 
diseases, as malarial, eruptive, typhus and typhoid fevers, 
croup, diphtheria, erysipelas, rheumatism, gout, peritonitis, 
meningitis, etc. ; 5. By lesions or prostration of the nervous 
system, especially when attended by diminished temperature 
and arterial tension, as from grief, fear, injury, pressure ; 
6. Pressure as from tumors, pregnancy, etc. ; 7. Cachexias, 
as from cancer, syphilis, scrofula, septicemia ; 8. Hydremia 
and ailments that disturb the vascular tension ; 9. Chorea, 
convulsions, exacerbations of febrile and other diseases ; 10. 
Diseases of genito-urinary organs, as Bright's disease, cystitis, 
hemorrhage, abscess, etc.; 11. Medicines, such as copaiba, cu- 
bebs, turpentine, some emetics and drastic cathartics, some 
anesthetics, coffee, many metallic salts, poisoning by hydrogen 
arsenide, carbon protoxide, carbon dioxide, phosphorus, iodine, 
iodoform, etc. 

The loss of organic material (albumin) disturbs nutrition, 
the blood becomes more aqueous ; it sometimes produces an 
anascarca which is the result of hydremia and of anuria. 

Albuminuria may be caused : 1. By an alteration in the 
renal transudation ; 2. By certain changes in the blood ; 3. By 
disturbance of the circulation. 

Renal Changes. Lesions of the dialysing portions of 
the kidney, especially of the glomerule; the epithelium of the 
convoluted tubules may also be essential in the production of 
albuminuria. These cells normally prevent the albumin from 
filtering through with the other elements of the plasma. Al- 
buminuria is also dependent upon renal lesions, sometimes 
primary as in nephritis, sometimes consecutive as in an alter- 
ation in the blood or a disturbance in the circulation. Acute 
nephritis, chronic nephritis, fatty or amyloid degeneration 
interfere with the process of dialysis. 



31 

Bacteria may exercise a traumatic action upon the renal 
epithelium and cause desquamation or degeneration, obstruct 
the vessels, modify blood pressure, or by the excretion of solu- 
ble products irritate the parts. 

Changes in the Blood. The dialysing membrane can- 
not stand with impunity any adulteration of the blood. The 
passage, in the kidney, of any such substance as biliary pig- 
ment in icterus ; glucose in diabetes ; poisons, such as alcohol, 
lead or mercury, or toxic gases, render the urine albuminous. 

Subcutaneous injection of solutions of extractives (leucin, 
tyrosin, creatinin, xanthin and hypoxanthin) cause degener- 
ation of the epithelium of the kidneys and albuminuria. The 
subcutaneous injection of tincture of cantharides causes, in a 
few minutes, the production of an albuminous exudate in the 
glomerules. Although the limit of saturation of the blood 
plasma by albumin may be unknown, it is none the less evi- 
dent that a superabundance of the substance (albumin) in the 
vessels causes albuminuria. 

The existence of a physiologic albuminuria is still doubt- 
ful in the domestic animals ; for Frohner, who has examined 
the urine of a number of healthy horses, has found only two 
cases in which albumin was present. In man it has been 
demonstrated to be due to severe muscular exercise, slight 
cold, and nitrogenous diet. 

Disturbance of Circulation. Too great a variation in 
blood pressure will cause albuminuria. 

Renal emboli, section of vaso-motor nerves of the kidney, 
medullary lesions, etc., cause albuminuria by causing an active 
congestion of the kidney. Venous stasis, organic affection of 
the heart, of the liver, presence of fetus, etc., cause albumin- 
ous urine. 

The urine may be less fluid. Bacteria may frequently 
cause thromboses, emboli, edemas, anemias, etc., from vaso- 
motor troubles changing simultaneously the filter, the liquids 
which filter, with regard to pressure and velocity. 

Albuminous urine is usually of light color and low specific 
gravity. It may occasionally be dark and dense, due to other 
ingredients, or to concentration. 

Under particular conditions of fatigue or disease albumin 
may appear in the urine. 



32 

Temporary albuminuria is sometimes induced by a cold 
bath, especially in persons prone to kidney disease, and it has 
been observed after excessive muscular exercise, as in the 
urine of soldiers after a prolonged march. 

Any cause which leads to an increased blood pressure in 
the kidneys tends to induce albuminuria, and many of the 
cases in which it is the result of disease may be traced to this 
cause. Albuminuria is a constant accompaniment of the 
nephritis following scarlet fever and may occur to a less extent 
in pneumonia, typhoid and diphtheria It may also occur in 
diabetes, and is then a highly unfavorable symptom. 

In every case the urine must be clear before testing, by 
filtering it carefully ; also take the specific gravity. In addi- 
tion to the suspected urines make control tests on the normal 
for comparison. 

Heat Test. Heat about 5 cc. of the urine to the 
boiling point in a test tube. Note the slightest turbidity. 
If present it will be due to albumin or earthy phos- 
phates. In horse urine the precipitate may be due to 
driving off C0 2 and precipitation of lime, etc., not phos- 
phates. Add slowly a few drops of acetic acid (or 
nitric). If due to the phosphates the urine becomes 
clear, if the turbidity remains it is albumin. Care must 
be taken, in the addition of the acid after boiling, to 
note the effect after each drop is added and to go on 
adding until there is no doubt that the urine is dis- 
tinctly acid. If only a trace of albumin is present and 
too much acid is added the albumin may be converted 
into acid albumin and remain in solution. Heat does 
not coagulate acid albumin. 

Another Heat Test. Fill a test tube about one- 
third full of water and boil it. Add a few drops of the 
suspected urine. If albumin is present a cloudiness or 
coagulum will appear, according to the amount of albu- 
min present. 

Heller's Cold Nitric Acid Test. Pour some of the 
urine gently upon the surface of some nitric acid in a 
test tube. A ring of white coagulum occurs at the 
junction of the two fluids. If the quantity of albumin 
is small, the coagulum may not occur for a few minutes. 



33 

A brown zone will frequently be seen at the point of 
contact due to the action of the acid upon the coloring- 
matters of the urine, but it does not give any turbidity 
unless .albumin be present. 

Millard-Robert's or Nitric Magnesian Test. The 
reagent is as follows: Nitric Acid, 1 part; Sat. Sol. 
Magnesium Sulphate, 5 parts. Use as in the preceding 
test. 

Picric Acid Test. (Johnson's). Pill the test tube 
half full of urine. Slightly incline the tube and gently 
pour down its side about 2 cc. of a saturated solution of 
picric acid, so that it may come in contact with the up- 
per layer of the urine. Place the tube in an upright 
position. A layer of coagulated albumin will appear at 
the line of junction. The coagulation of albumin takes 
place at once and is thus not easily mistaken for precip- 
itated urates, which require some time for their precip- 
itation and disappear on the application of heat. 

Ferrocyanide Test. To 2 cc. of acetic acid in a 
test tube add 4 cc. of a 5% solution of potassium ferro- 
cyanide. Mix them and add 10 cc. of urine. A precip- 
itate will appear if albumin be present. No heat is 
required. 

A test equally as good is that proposed by Zouchloss 
in which potassium sulphocyanide is substituted for the 
ferrocyanide. 
Sugar in the Urine. Dextrose or glucose exists in the 
blood from 0.8 to 1.25 parts per 1000. When a greater amount 
than 3 parts per 1000 exists, the excess is excreted through 
the kidneys. It is maintained by many that a trace of dex- 
trose is normally present in the urine and may appear in 
slightly larger quantities transitorily without pathologic 
significance. The presence of small quantities of sugar in 
the urine is desig-nated as glycosuria ; in larger quantities it 
is known as diabetes mellitus. The former condition if habi- 
tual, is unnatural, and may terminate in the latter. 

These diseased conditions do not necessarily point to dis- 
eases of the kidney or urinary organs, but rather to the liver. 
The kidney, in ridding itself of this product (dextrose), becomes 
irritated, and this irritation extends down the entire canal, 
and we thus have a real polyuria produced. 



34 

Sugar, in sufficient quantity to react to ordinary tests, is 
found in the urine physiologically during" pregnancy and lac- 
tation ; in infants under two months old ; in old persons living 
largely upon starchy and saccharine food. Pathologically in 
diabetes mellitus ; in impeded respiration from pulmonary 
diseases ; in impeded hepatic circulation (functional and or- 
ganic diseases of the liver); in diseases of the central nervous 
system (general paresis, epilepsy, dementia, puncture of the 
fourth ventricle); in intermittent and typhoid fevers, by the 
action of certain poisons, as carbon monoxide, arsenic, chloro- 
form and curare ; in abnormally stout persons. 

The persistent excretion of easily recognizable quantities 
of sugar constitutes diabetes. The quantity of urine is often 
enormously increased, as much as 10000 cc. being passed in 24 
hours, by man. The specific gravity is high, varying from 
1025 to 1050. The color is usually pale, from the dilution — 
not diminution — of the urinary pigments. 

The presence of albumin interferes with the tests for sugar 
and must, in all cases, be removed by the addition of acetic 
acid and heat. The urine, after being filtered, may then be 
used for the sugar tests. 

The particular property of glucose, which is utilized for 
its detection, is its action as a reducing agent — its disposition 
to absorb oxygen. In this property it differs strikingly from 
saccharose or common cane sugar. 

Principle of the Copper Tests. If a little copper sulphate 
and an excess of a solution of caustic potash be added to a 
solution of glucose, a clear blue solution results. Without the 
glucose, the alkali would precipitate the pale blue cupric hy- 
drate, Cu0 2 H 2 ; and if the mixture were boiled this blue pre- 
cipitate would be reduced to a black precipitate of Cupric 
Oxide, CuO,. The clear blue solution containing glucose, 
however, when boiled, changes from transparent blue to opaque 
yellow, and speedily deposits a yellow, ultimately red, precip- 
itate of cuprous oxide CuO. When the quantity of sugar is 
large the change is immediate. When small the reaction takes 
a few minutes for its completion. 

Pehling's Solution. Solution A. 34.64 grams of pure 
crystalline copper sulphate are powdered and dissolved in 500 
cc. of distilled water. Solution B. Sodio-potassium tartrate 



35 

(Rochelle Salts) 173 grams. Pure caustic potash 125 grams. 
Add enough distilled water to make 500 cc. When using take 
equal parts of Solutions A and B. (If diluted with 5-10 vols. 
of water the test is said to be more sensitive). 

Place some Fehling's Solution in a test tube and 
boil it. If no yellow discoloration takes place it is in 
good condition. Add a few drops of the suspected urine 
and boil. If the mixture suddenly turns to an opaque 
yellow or red color, the presence of a reducing sugar is 
indicated. Normal horse urine usually changes the 
color of the copper solution to a black or dark tint, but 
this does not indicate sugar. 

Trommer's Test. To 4 or 5 cc. of urine, in a test 
tube, add one-half its volume of 20% caustic potash and 
2 or 3 drops of a solution of copper sulphate (1-10). 
Heat to the boiling point. If sugar be present a yel- 
lowish or reddish precipitate is thrown down, the sugar 
having reduced the cupric hydrate to cuprous oxide. 

Bismuth Test. Put equal quantities of urine and 
20% solution of caustic potash in a test tube and add a 
pinch of subnitrate of bismuth. Boil the mixture and 
if glucose be present the powder turns black. Albumin 
and sulphur also reduce bismuth and must be removed 
if the test is to be reliable. Bismuth is more or less 
blackened with normal horse urine. 

A better form of the bismuth test is as follows : A 
solution is made of bismuth subnitrate 2 grams ; Ro- 
chelle salts 4 grams ; sodium hydroxide 8 grams ; and 
distilled water to 100 cc. The urine is heated to boiling 
and a few drops of this alkaline solution of bismuth 
added, and on continuing the boiling, if sugar be pres- 
ent, the mixture turns black. The test is delicate, as 
little as 0.025% of glucose can be detected. 

Phenylhydrazine Test. (Modified from Kowarsky). 
To 5 drops of pure phenylhydrazine and 10 drops of 
glacial acetic acid in a test tube is added 1 cc. of a 10% 
solution of sodium chloride. After shaking the mixture, 
3 cc. of the urine are added and the test tube heated for 
two minutes or longer. The fluid is then allowed to 
cool slowly. If the sugar contents exceed 0.5% the pre- 



36 

cipitate of glucosazone takes place in about two minutes. 
Small quantities of albumin do not hinder the reaction, 
but are precipitated by boiling. After an hour or more 
examine the precipitate microscopically. A 10% solu- 
tion of sodium hydroxide has been recommended as a 
substitute for the sodium chloride solution and greater 
delicacy is claimed for the reaction. 

The glucosazone crystals will have the form typical 
of skeins of wool, which will generally be double, 
whereas other crystals precipitated, such as those of 
glycuronic acid, are irregularly formed. As little as 
0,005% (one-fortieth grain per ounce) of glucose can be 
detected in urine by the phenylhydrazine reaction. 

Uric acid, urea, xanthine, and creatinin in no way 
simulate the reaction of glucose with phenylhydrazine. 
The only bodies which can offer confusion are glycur- 
onic acid and its derivatives. 

Agostini's Test. Urine, 5 drops; 0.5% solution 
gold chloride, 5 drops ; 20% solution caustic potash, 3 
drops. Heat gently. Contact with glucose gives red 
color. 

Fermentation Test. Robert's Differential Density 
Method. Take the specific gravity of the urine before 
adding the yeast and record it. Mix well 2 fluid ounces 
(60 cc.) of urine with % cake of compressed yeast in a 
bottle. Set aside for 24 hours in a moderately warm 
place. After the fermentation filter and take the spe- 
cific gravity again and subtract from that taken before. 
Each degree of the remainder represents one grain of 
glucose to the fluid ounce. Multiply by 0.219 to get the 
percentage. Thus : Specific gravity before fermenta- 
tion, 1035 ; specific gravity after fermentation, 1015. 
1035 — 1015=20 degrees of density lost, or 20 grains of 
sugar to the fluid ounce. This test is conclusive as to 
the presence of sugar, though it is not absolutely accur- 
ate as to quantity. 



37 



VI. 



Bile in the Urine. In a number of pathologic condi- 
tions the elements of the bile are excreted in the urine. The 
bile pigments, bilirubin and biliverdin, may occur along with 
the bile salts, sodium glycocholate and taurocholate, or the 
bile salts alone may be present. 

Urine containing the bile pigments is colored a yellow 
brown or brownish green. It forms an intense yellow froth 
on agitation. It stains paper or linen a permanent yellow. 
The bile pigments are found in jaundice, functional disorders 
of the liver (acute and chronic biliousness); organic diseases 
of the liver apart from jaundice (carcinoma, amyloid disease, 
cirrhosis) ; diseases of the spleen ; fever ; hemolytic diseases 
(anemia, leucocythemia and scurvy). 

Gmelin's Test. (Nitric acid containing nitrous 
acid). Place a few drops of the suspected urine in a 
white porcelain dish and near them a few drops of the 
impure nitric acid ; let the fluids run together and the 
usual play of colors is observed. 

Put some urine in a test tube and carefully pour in 
some of the yellow impure nitric acid, until it forms a 
stratum at the bottom. If the bile pigments are present 
at the line of junction of the fluids, a play of colors 
takes place — from above downwards — green, blue, violet 
or dirty red, and yellow. Nearly all urines give a play 
of colors, but green is the necessary and characteristic 
color to prove the presence of the bile pigments. 

Pettenkofer's Test for Bile Acids. Take about 2 cc. of 
suspected urine in a test tube and add 4 drops of a 10% 
solution of the cane sugar. Add strong sulphuric acid, 
drop by drop, cooling the tube in a dish of cold water 
immediately after adding the acid. Not more than 2cc. 
of the acid should be used. Too much heat causes car- 
bonization of the sugar and the test is ruined. If bile 
acids are present, the fluid at first becomes opaque, then 
clear and successively brown, red and purple. It may 
require an hour or more to accomplish the test. This 
reaction depends upon the production of furfurol by the 



38 

destruction of the sugar when the sulphuric acid is 
used. Furfurol in turn combines with cholalic acid, 
formed by the action of the sulphuric acid on the bile 
acids, giving - the color. 

Pettenkofer's test may also be quite satisfactorily 
performed more quickly by putting a little of the sus- 
pected urine in a porcelain capsule, adding a few drops 
of a solution of cane sugar and then a few drops of 
strong sulphuric acid. Keeping the mixture cool to pre- 
vent carbonization. 

Udranszky has modified Pettenkofer's test by using 
furfurol directly instead of waiting for its formation by 
the action of the sulphuric acid upon the sugar. His 
procedure is as follows : One cubic centimeter of the 
urine is treated with one drop of a 0.1% solution of fur- 
furol and slowly superposed upon 1 cc. of sulphuric acid, 
care being taken to prevent too great heating of the 
mixture. A purple color appears at the plane of con- 
tact, that gradually extends upward into the superposed 
solution ; on standing, the color turns bluish. In alco- 
holic solution a green flourescence is seen. The pigment 
gives a typical spectrum. 

Hay's Test. Use two test tubes ; in one place a 
little normal urine and in the other some of the suspected 
urine. Leave the test tubes in the rack in order to pre- 
vent agitation. Drop a little finely powdered sulphur 
upon the surface of the urines. If bile or biliary acids 
are present in the suspected urine the sulphur will sink 
at once to the bottom of the tube, while in the normal 
urine it will remain upon the surface or but a slight 
amount may sink, if the tubes are not agitated. 

Chloroform as a test for bile is quite satisfactory. 
Agitate a few drops of chloroform with the suspected 
urine in a test tube If bile be present the chloroform 
becomes turbid and acquires a yellowish hue, the depth 
of which is in proportion to the amount of bile present. 
To some of the suspected urine add a little bromine 
water. If bile is present a green ring should appear. 



39 

Blood. Blood is sometimes a constituent of the urine in 
disease. It may occur in two forms : 1. As hematuria, when 
the blood coloring matter is present in the urine in combination 
with blood corpuscles. 2. As hemoglobinuria, when no blood 
corpuscles are present and the blood pigment or hemoglobin 
is in solution in the urine. 

Hematuria may have its source (1) in the kidneys due to 
injuries; acute nephritis; acute acerbation of chronic nephritis; 
diseases of renal vessels (embolism, thrombosis, aneurism, 
stasis); amyloid kidney (very rarely); infective fevers (small- 
pox, scarlatina, typhoid fever, etc.); certain blood diseases 
(scurvy, purpura, hemophilia) ; parasitic diseases (echino- 
coccus). 2. The renal pelvis and ureters due to renal calculi ; 
tuberculosis ; rupture of neighboring abscesses ; parasites. 
3. The bladder due to calculi ; cancer and other tumors ; diph- 
theritic cystitis ; varicose veins ; injuries. 4. The urethra due 
to injury (catheterization, impaction of calculi, etc.). 5. Ex- 
traneous discharges as the menstrual flow, etc. 

Hemoglobinuria has been observed in severe infectious 
diseases (typhoid fever, scarlatina, etc.); in conditions of 
blood dissolution (scurvy, purpura, etc.); in skin burns, sun- 
stroke, etc. 

Heller's Blood Test is made by adding a little caus- 
tic soda solution to some urine in a test tube and heating. 
The precipitated phosphates are colored reddish brown 
and fall in a thick cloud to the bottom of the tube. 

Almen's Test. Add a few drops of freshly made 
tincture of guaiacum to the suspected urine. Shake 
well and add a few drops of hydrogen dioxide or of old 
oil of turpentine, and the mixture will turn greenish 
blue. The hemoglobin will change the color of the 
precipitate to blue. The test will not respond to a 
small quantity of blood. 

Hemin Test. If some blood or sediment supposed 
to contain blood be heated carefully with some glacial 
acetic acid and a trace of sodium chloride, and then 
slowly evoporated in the air, brownish yellow rhombic 
crystals of hemin are found. 

The spectroscope and microscope are also used in 
blood tests. 



40 

Melanine. la certain cases of melanosis this pigment 
appears in the urine, which, when emitted is clear, but grad- 
ually becomes of a deep brown, or even black color. 

Ordinarily melanine exists in solution in the urine, but 
sometimes in the form of brownish or black sediment, recog- 
nizable by microscopic examination. Melanine may possess a 
diagnostic significance when the melanosis is beyond the reach 
of examination by eye or touch. 

It may disappear from the urine when the disease is 
arrested, or it may remain stationary. Oxidizing agents, such 
as chromic acid and fuming nitric acid, transform the principle 
melanine, causing gradually with the first and immediately 
with the second a black coloration. According to Zeller, the 
most delicate test for melanine is bromine water. With mel- 
anine it gives at first a yellow precipitate, which gradually 
blackens. Urobilin gives a yellow precipitate with the same 
reagent, but it does not blacken. 

The practical significance of this condition (melanuria) is 
greatly limited by the fact that the urine may contain a large 
quantity of melanine in wasting diseases, whilst that derived 
from individuals suffering from melanotic cancer or sarcoma 
may be entirely free from it. Senator has recently confirmed 
this view by a series of clinical observations. Nevertheless, 
as an adjunct in diagnosis, the tests given are of undoubted 
utility. 



VII. 
QUANTITATIVE ANALYSIS. 

Centrifugal Method. The centrifuge as ordinarily used 
gives the bulk percentage of the constituents, this percentage 
being arrived at arbitrarily as a result of numerous determin- 
ations upon normal urine. Knowing the normal bulk percent- 
age it is not difficult to determine an abnormal or pathologic 
amount of a substance as indicated by its excess or deficiency, 
and the method is therefore a convenient and expeditious one for 
clinical purposes. To a beginner, however, the method as com- 




41 

monly employed, is misleading' in that 
the bulk percentage and the tine or actual 
percentages are widely different. Take 
for example the phosphates of the urine. 
The normal bulk percentage as given 
by the centrifuge is 8% for man ; l c /o 
or less for the horse. Whereas, the 
real amount of phosphates present as 
P 2 5 in the urine of man for the 
whole 24 hours (1500 cc. of urine) is 
only about 3.5 grams, or in 1000 cc. 
about 2.5 grams, or a true percent- 
age of 0.25%. The true percentage 
may be calculated from the centrifuge 
by giving to each 0.1 cc. of precipitate 
a determined value with reference to 
the amount of P 2 5 present. 



Fig. 9. 
Hand Centrifuge. 




Fig. 10. Aluminum Tubes. 



Percentage 
Tube. 



Sedimentation 
Tube. 



Each centrifuge tube is graduated to 15 cc. The first 
10 cc. are divided so that each cubic centimeter is divided into 
10 parts, each part representing 0.1 cc. The remaining 5 cc. 
are for holding the test reagrents which are added to the 10 cc. 
of urine. The first cubic centimeter because of the tapering 
end, will admit of finer graduation than the remainder of the 
tube. The first half of the cubic centimeter is divided into 



42 

■fa cc. (or .025 cc. ) in order that small amounts of precipitate 
may be accurately read ; the second half of the cubic centimeter 
is divided into 2V cc - (or.OScc); while the remaining- 9 cc. 
are each divided into fa cc. (or 0.1 cc.) as above stated. It is 
convenient to take the fa cc. as the unit in the calculation of 
the true percentage. By taking - the average of a great num- 
ber of control tests with the burette it has been found that 
fa cc. of the precipitate, in the case of the phosphates, repre- 
sents 0.1313 gram of P 2 5 in 1000 cc. of urine. 

If albumin is present, remove it by adding a little acetic 
acid and applying heat. Filter and test the filtrate for the 
inorganic constituents. 

With the horse urine care must be exercised in adding the 
reagents. The large amount of carbonates present cause con- 
siderable effervescence, from the liberation of the C0 2 , and 
some of the urine is likely to flow over the tube. Add a little 
of the reagent and when the effervescence has ceased add a 
little more until the required amount has been used. 

Phosphates. Fill the graduated tube to the 10 cc. 
mark with the urine to be tested. Add 1 cc. of glacial 
acetic acid and 4 cc. of 5 l /o uranium nitrate solution to 
reach the 15 cc. mark. Invert the tube several times 
and revolve in the centrifuge for three minutes. If, 
after revolving three minutes at 1000 revolutions per 
minute, the precipitate comes up to the eighth 0.1 cc. 
line of the tube (0.8 cc.) multiply 0.1313 by 8 for the 
product, which is 1.0504 grams of P 2 5 in 1000 cc. 
If the total amount of urine for the 24 hours is 1400 cc, 
the amount of P 2 5 present in this quantity can easily 
be calculated by the following proportion : 1.0504 gm. 
P 2 O s : 1000 cc. :: X : 1400. In which the value of X is 
found to be 1.470 grams of P 2 5 in 24 hours. Make 
the same determination with the urine of the horse. 

Chlorides. Fill the graduated tube to the 10 cc. 
mark with the urine. Add 15 drops or 1 cc. of nitric 
acid to prevent precipitation of the phosphates. Then 
fill to the 15 cc. mark with the silver nitrate solution 
(1 to 8). Invert the tube several times to thoroughly mix 
the reagents. Revolve in the centrifuge for intervals 



43 

of three minutes until the precipitate no longer settles. 
The average amount of precipitate for man is from 1 cc. 
to 1.2 cc; for the horse about 1 cc. The value of each 
0.1 cc. is 1.541 of a gram of NaCl per 1000. 

Sulphates. These are estimated as insoluble salts 
of barium. Fill the graduated tube to 10 cc. mark with 
fresh urine and add 5 cc. barium chloride solution, (4 
parts barium chloride, 1 part hydrochloric acid, 16 parts 
distilled water). 

The amount of precipitate for man and horse is 
about .075 cc. normally. The value of each 0.1 precipi- 
tate is 2 grams of SO s for both man and horse. 

In the above tests write down in your notes the 
amount of each constituent for the 24 hours considering 
1250 cc. as the total amount of urine passed. Use the 
hand, water and electric centrifuges in regular order, 
stating after each result, which centrifuge was used. 
Quantitative Estimation of Uric Acid. This estima- 
tion is difficult and time-consuming and is generally regarded 
as inexpedient for clinical purposes. The following methods 
are described because they are more suitable for clinical work 
although less accurate than the more elaborate methods of 
Salkowski-Ludwig, Hopkins and others. 

Ruhemann's Uricometer for the rapid estimation of uric 
acid consists of a graduated tube with a glass stopper. It is 
a colorimetric method, in which iodine with carbon bisulphide 
serve as indicators. 

The following directions accompany each apparatus : 
Fill the glass tube to the lowest mark S with carbon bisul- 
phide. (It is not necessary for the tube to be quite dry, but 
there must be no drop of liquid at the bottom). 

The lowest part of the convexity (double meniscus) has to 
be even with mark 5, see diagram. 

Add a solution consisting of 1.5 gm. iodine ; 1.5 gm. potas- 
sium iodide; 15 gm. absolute alcohol and 185 gm. distilled 
water. 

Fill up so that the base of the upper arch of the double 
meniscus is on a level with mark/ as shown in the illustration. 
Then add the urine to be examined, which has to be at a 
temperature of 18° Centigrade, to the mark 2.45 (2.6 ccm). 



44 



4.0- f: 



Fig. ll. 
Ruhemann's 
Uricometer. 



Close the tube with glass stopper a n d 
shake well when the carbon bisulphide will 
become a dark copper brown color. 

After adding more urine under continued 
Strong shaking, the carbon bisulphide will 
absorb all free iodine and the mixture will 
look like urine. 

Slowly adding more urine will change 
the yellow foam, created by the shaking, 
into white foam. 

The color of the carbon bisulphide will 
turn pink after a while. 

Should this color remain the same after 
the apparatus has been shaken repeatedly 
and turned upside down, add another drop 
of urine and keep up the same procedure un- 
til only a slightly reddish coloration of the 
carbon bisulphide remains. 

Now shake again vigorously and the car- 
bon bisulphide will turn porcelain white and 
the urine will look like cloudy whey. 

To recapitulate : — The adding of urine 
has to be stopped as soon as the carbon bi- 
sulphide shows only a slightly reddish tint, 
because this will disappear entirely after re- 
peated shakings. The test is finished when the 
indicator appears snow-white, a sign thai all iodine 
has been neutralised by the urine. 

To get rid of the remaining foam move 
the tube a few times slowly to a horizontal 
position, then open the stopper a little, to 
allow all liquid to settle in the tube. 

The proportion of uric acid is then read 
off the upper scale (per thousand of urine). 

The percentage is obtained by putting 
a in front. If the upper meniscus line 
of the urine is between any of the 0.1 ccm. 
marks, the upper number should be read. 

Should the urine contain less uric acid 
than the apparatus will in this way indicate, 



45 

add the iodine solution to the mark half way between 5 and J 
and read after each reaction the half values. 

The vessel in which the urine is to be kept, must not be 
cleaned with soda. 

If the urine shows an acid reaction, it can be used at once, 
but if it should be alkaline, it has to be made acid by adding 
diluted acetic acid. Cloudiness is of no importance. 

If the urine contains a considerable sediment of sodium 
urate it should be well shaken. 

Strong colorations of the urine do not affect the action of 
carbon bisulphide. 

Traces of sugar and albumin do not disturb the result. 

If there is a very large percentage of albumin or traces of 
blood or pus, these pathologic substances have to be coagu- 
lated by boiling and the urine filtered. 

(The apparatus may be purchased of Eimer & Amend, 

New York). 

Cook's Method of Estimating Uric Acid by the Centrifuge. Place in 
the graduated tube 10 cc. of urine ; add to this 1 gm. of sodium car- 
bonate and l cc. of ammonium hydrate. Shake until the sodium car- 
bonate is dissolved ; this precipitates the earthy phosphates. Separate 
this precipitate with the centrifugal machine and decant the supernatant 
urine into another graduated tube. It will be found that the earthy 
phosphates are readily separated and adhere to the bottom of the tube ; 
this allows the clear urine to be readily poured off into another tube. 
To the clear urine now free from phosphates add 2 cc. of ammonium 
hydrate and 2 cc. of ammonio-silver solution (made by dissolving 5 gm. 
of silver nitrate in 100 cc. of water and adding ammonia until the solu- 
tion becomes clear). The addition of the silver solution causes the uric 
acid to be precipitated as the urate of silver, a translucent, shiny sub- 
stance. Separate this precipitate with the centrifugal machine and pour 
off the supernatant urine. Add to the ppt. an excess of ammonium 
hydrate, at least 15 cc. and mix thoroughly. By this last addition any 
of the chlorides that may have been precipitated are redissolved leaving 
only a pale urate of silver. Lastly precipitate this urate of silver until 
the lowest reading is to be had. Each 0.1 cc. as marked on the gradu- 
ated tube indicates 0.001175 gram of uric acid in 10 cc. of urine. The 
amount of uric acid for the 24 hours can be calculated by the following 
formula, supposing that 1250 cc. was the amount of the urine passed. 

0.001175X5 1 ' 2S0=0 . 7343gm . 



10 

Estimation of Uric Acid by Weight. 20 cc. of hydrochloric acid are 
added to 200 cc. of urine and the mixture set aside for 24 hours. Uric 
acid crystals form and collect on the sides of the vessel. This may be 
collected on a weighed filter and washed thoroughly with water. The 
filter and uric acid are dried and weighed until there is no further loss 
of weight. The weight of the filter paper is deducted and the result 
gives the amount of uric acid in 200 cc. of urine, from which the amount 
in 24 hours can be calculated. 



46 




Urea. Doremus Ureotneter Test. Fill the 
long- arm and bend of the ureotneter with the 
hypobromite solution. (Hypobromite of sodium 
is prepared by mixing- 2 cc. of bromine with 
23 cc. of a solution of caustic soda, 40 grams to 
100 cc. of distilled water. To this mixture 
add an equal volume of distilled water). With 
a thoroughly washed pipette draw up exactly 
1 cc. of urine and pass the pipette through the 
bulb of the ureometer as far as it will go in 
the bend. Compress the bulb of the pipette 
gently and steadily. The urine will rise 
through the hypobromite, and the urea in- 
stantly decompose, giving off nitrogen gas. 
Withdraw the pipette after the urine has been 
expelled, taking care not to press the bulb hard 
enough to drive the air out with the urine, and 
read the volume of gas, after allowing the 



r~\ 




froth to subside. 

Each division mark on the 
ureometer indicates 0.001 gram 
of urea in 1 cc. of urine. The 
quantity of urea voided in 24 
hours is ascertained by multi- 
plying the result of the test by 
the number of cc. of urine passed 
during that period. 

The C0 2 resulting from the 
decomposition of the urea is ab- 
sorbed by the excess of soda in 
the hypobromite solution, and 
nitrogen is evolved. 37.1 cc. of 
moist nitrogen gas measured 
under the ordinary conditions 
of experiment may be taken 
to represent 0.1 gram of urea. 
1 gram of urea corresponds to 
13.72 grams of muscular tissue. 



FIG. 13. Hind's modification of 
Doremus Ureometer. 



47 



Albumin. Esbach's Method. Fill the 
tube with the suspected urine to the letter U, 
then add the reagent to the letter R. (The 
reagent is made by mixing 10 grams of picric 
acid and 20 grams of citric acid and adding 
enough water to make 1 liter. It is said that 
acetic acid may be substituted for the citric 
with as good results). Invert the tube a 
number of times so that the contents may be 
thoroughly mixed. Close the tube tightly 
with the rubber stopper and set aside for 24 
hours, after which the amount of dried albu- 
min in one liter of urine can be read in grams 
on the tube. The percentage is obtained by 

dividing- by ten. Thus, if the coagulum FlG- 14 - 

Esbsch's 
stands at 3, the urine contains three parts of Albumin- 
albumin per thousand, or 0.3%. If the albu- ometer. 
min is very abundant (above four) the urine should be 
diluted to obtain an accurate result. Less than 0.5 
per thousand of albumin cannot be accurately estimated 
by this method. 

Caution. The coagulable substance found in the 
urine of Bright's disease is a mixture of " serum- 
albumin " and " serum-globulin " or "para-globulin." 
Saturation of the urine with crystallized magnesium 
sulphate precipitates the serum-globulin, leaving the 
serum-albumin in solution. 

Centrifuge Test. Fill the graduated tube with urine 
up to the 10 cc. mark ; add 5 cc. of Esbach's reagent. 
Invert the tube several times in order to thoroughly mix 
the fluids. Place the tube in the centrifuge and revolve 
for three minutes. Read off the amount of the precipi- 
tate ; each 0.1 cc. is the equivalent of 0.225 of a gram 
of dry albumin per 1000 cc. of urine. 

Ferrocyanide Test with Centrifuge. The percentage 
tube is filled to the 10 cc. mark with the urine to be 
tested. Add 1 cc. of acetic acid and 4 cc. of a 5% solu- 
tion of potassium ferrocyanide. Proceed as in the pre- 
ceding test. Each 0.1 cc. of precipitate is the equivalent 
of 0.222 gram of dry albumin per 1000 cc. of urine. 




48 

Sugar. Einhorn's Fermentation Saccharometer. 
Determine the specific gravity. Add 10 cc. of the sus- 
pected urine to 90 cc. of distilled water. Put in a flask, 
add about one gram of compressed yeast and agitate 
thoroughly. Pour 10 cc. of the mixture into the 
bulb of the saccharometer and, by inclining the 
apparatus, the fluid will displace the air in the 
cylinder and remain there by atmospheric press- 
ure. Be sure that no air bubbles remain in the 
cylinder. It is always well to test a normal urine 
at the same time and in the same way as a con- 
trol. The mixture of normal urine with yeast 
will, on the following day, have only a small 
bubble on the top of the cylinder. If, in the sus- 
Einhorn's pected urine, there is also present at the top of 
Saccharo- the cylinder a small bubble, no sugar is present ; 
but if there*is a much larger volume of gas (C0 2 ) 
it is certain that the urine contains sugar. The appar- 
atus should remain in a moderately warm place. As a 
result of fermentation the sugar is broken up into alco- 
hol and C0 2 . The changed level of the fluid in the 
cylinder shows that the reaction has taken place and 
indicated by the numbers the approximate quantity of 
sugar present. The scale on the tube is empirical and 
indicates directly the percentage of sugar in the urine. 

Shieb's Test for Sugar in the Urine. 
Solution No. 1. 

Ammonium Sulphate (purest) 
Copper Sulphate (purest) 
Distilled Water 
Solution No. 2. 

Caustic Potash C. P. 

Distilled Water 

Dissolve and when cool, add 

Glycerine 

Ammonia water 0.960 sp. gr. 

Add No. 1 to No. 2 and dilute the whole to 500 cc. 
with distilled water. Stopper securely and shake till 
thoroughly mixed. 



1.2 


grams. 


2.6 


grams. 


50. 


cc. 


20 


grams. 


50 


cc. 


50 


cc. 


300 


cc. 



49 

Heat one dram of this solution in a test tube to boil- 
ing-. Add the urine drop by drop, at slow intervals, 
boiling- after each addition until the blue color has been 
discharged and the fluid has a lig-ht amber color or is 
colorless. 

If the solution is decolorized by 3 minims of urine 
it contains 9 to 10 grains of sugar per oz. 

If the solution is decolorized by 4 minims of urine 
it contains 7 to 8 grains of sug-ar per oz. 

If the solution is decolorized by 5 minims of urine 
it contains 5 to 6 grains of sug-ar per oz. 

If the solution is decolorized by 6 minims of urine 
it contains 4 grains of sug-ar per oz. 

If the solution is decolorized by 7 minims of urine 
it contains 3 grains of sug-ar per oz. 

If the solution is decolorized by 9 minims of urine 
it contains 2 grains of sugar per oz. 

If the solution is decolorized by 10 to 17 minims of 
urine it contains 1 grain of sug-ar per oz. 

If the urine contains more than 10 grains of sugar 
to the ounce it must be diluted with an equal quantity 
of water, and the number of grains per ounce multiplied 
by two. 

A very similar preparation, accurately compounded, 
is on the market under the name of Whitney's Reagent. 
It is for sale by the Norwood Chemical Co., 105 W. 40th 
St., New York. 

The following table gives the amounts of sugar in 
analytical testing with Whitney's Reagent : 

If reduced by It contains to the oz. Percentage. 



1 minim 


16 grains or more 


3.33 


2 minims 


8 grains 


1.67 


3 " 


5.33 


grains 


1.11 


4 " 


4 


(< 


0.83 


5 " 


3.20 


<i 


0.67 


6 " 


2.67 


(< 


0.56 


7 " 


2.29 


ii 


0.48 


8 


2 


i< 


0.42 


9 " 


1.78 


(C 


0.37 


10 " 


1.60 


(( 


0.33 




(1 


(( 


0.21) 



50 

Ehrlich's Diazo-reaciion. This test has been recom- 
mended for the diagnosis of typhoid fever in man. It is 
stated that the reaction will also occur in some other 
diseases, but in spite of this fact the test is of value as 
an aid in the diagnosis. Two solutions are prepared as 
follows : 

1. Sulphanilic acid 2 gms. 
Hydrochloric acid 50 cc. 
Distilled water 1000 cc. 

2. A. 0.5% solution of sodium nitrite. 

In performing the test, 50 parts of No. 1 and 1 part 
of No. 2 are mixed, and equal parts of this mixture and 
of the urine in a test tube are rendered strongly alka- 
line with ammonia. If the reaction be positive, the 
solution assumes a carmine-red color, which on shaking 
must also appear in the foam. Upon standing 24 hours 
a greenish precipitate is formed. The test must not be 
considered positive unless a distinct coloration extends 
to and includes the foam on shaking. 



VIII. 



VOLUMETRIC METHODS. 



Chlorides. Mohr's method. Principle : If silver nitrate 
be added to a solution containing sodium chloride, neutral po- 
tassium chromate, and an alkaline phosphate, the chloride is 
first precipitated, then the chromate, and lastly, the phosphate. 
The formation of the red silver chromate indicates the com- 
plete precipitation of the chloride. 

Solutions required : 

1. Standard solution of silver nitrate : 

Fused silver nitrate 29.075 grams. 
Distilled water to make 1000 cc. 

2. Saturated solution neutral potassium chromate: 

Neutral potassium chromate 10 grams. 
Distilled water to make 100 cc. 



51 

Process, (a) The urine should not be high colored, 
and should be free from albumin or excess of uric acid 
or mucus. Dilute 10 cc. of the urine with lOOcc. of dis- 
tilled water. Fill a burette with the silver solution to 
the zero mark. Drop it slowly into the urine, stir it 
well and occasionally carry a drop of the mixture so as 
to come in contact with a little of the chromate solution 
in an evaporating- dish Test in this way until the first 
trace of orange color appears in the chromate solution. 
Make sure that the precipitation of the chlorides is 
complete by adding another drop of the silver solution 
from the burette. Read off the amount of silver solu- 
tion used and calculate the result. 

Example : 

Quantity of urine in 24 hours, 1250 cc. 
Silver solution used, - 7.5 cc. 

1 cc. silver solution equals .01 gram NaCl 
.01 X 7.5 



10 



X 1250=9.375 grams. 



Make two or three determinations and take the 
average for your final result. Do the same in the phos- 
phate and sulphate tests. 

If the urine of the horse is very dark colored, it may 
be filtered through animal charcoal to make it light 
colored Some of the chlorides may be held by the char- 
coal and thus diminish the amount in the urine tested ; 
or the urine may be diluted with an equal volume of 
distilled water and the result multiplied by two. 

A more accurate method is the following: (b) If 
the urine is high colored, and contains albumin, or excess 
of uric acid and mucus, they must be removed, To do 
this, measure 10 cc. of the urine into a platinum capsule, 
add 2 grams of pure potassium nitrate, evaporate to 
dryness, and ignite at a dull red heat to destroy organic 
matter. When cool, treat the residue with hot water 
and filter. Acidulate the filtrate with dilute nitric acid, 
neutralize with carbonate of lime and proceed as in (a). 



52 

Estimation of Phosphoric Acid. (Estimated as P 2 5 ). By 
uranium acetate or nitrate. This method is based upon the 
fact that when a solution of acetate or nitrate of uranium is 
added to a solution containing soluble phosphates, sodic acetate 
and free acetic acid, all the phosphoric acid will be precipi- 
tated as phosphate of uranium. This precipitate is of a light 
yellow color, insoluble in acetic, but soluble in hydrochloric 
acid. The point of completion of the phosphoric reaction may 
be ascertained by placing with a glass rod a drop of the yellow 
mixture in contact with a drop of potassium ferrocyanide solu- 
tion upon a white plate or filter paper. As soon as there is 
the slightest excess of the uranium solution after the phos- 
phates have been satisfied, a brown precipitate will result in 
the mixture due to formation of ferrocyanide of uranium. 
The cochineal solution, noted below, serves as a better indi- 
cator for the horse urine than the ferrocyanide. The follow- 
ing solutions are used : 

1. Solution of Potassium Ferrocyanide. 

Potassium ferrocyanide, - 5 grams. 
Distilled water - 100 cc. 

(Or a solution of cochineal prepared by boiling 40 grams 
of cochineal in 800 cc. of water. When cool add 
200 cc. of alcohol and filter). 

2. Solution of Sodium Acetate. 

Sodium acetate, - 100 grams. 

Acetic acid, - - 100 cc. 

Distilled water to make 1000 cc. 

3. Standard Solution of Disodic Hydric Phosphate, made 
by dissolving 10.0845 grams of the crystallized salt in water 
and diluting to 1 liter. Each cc. of this solution contains .002 
of a gram of P 2 0, 3 . In 5 cc. there is 0.1 gram of P 2 O s . 

4. Uranium Acetate. Since this cannot be obtained 
sufficiently pure to be weighed out and used directly we make 
a solution of it of indefinite strength and standardize it with 
the other solutions. 

It has been found best to make the solution of uranic ace- 
tate of such a strength that each cc. will precipitate .005 of a 
gram of P 2 5 . Consequently every 2 cc. of the uranium ace- 
tate should be made equal to every 5 cc of the sodic phosphate, 
or upon adding 20 cc. of the uranium acetate to 50 cc. of the 



53 

sodium phosphate and then touching- the plate or paper which 
has been moistened with the cochineal solution with the drop 
of the mixture, we should just get the green color. 

Put 50 cc. of the sodium phosphate with 5 cc. of the sodium 
acetate solution into a beaker. To this add slowly from the 
burette, the uranium acetate, testing occasionally, for the 
color on the paper. Suppose that on the addition of 8cc. from 
the burette, the color is obtained, then 8 cc. of the uranium 
acetate are as strong as 20 should be, and for every 8 cc. of the 
uranium solution that we have, 12 cc. of water should be added. 
If it should require more than 20 cc. to produce the color, the 
uranium solution must be concentrated by evaporation or more 
of the solid salt added. The solution has now been graduated 
or standardized. 

Application. 50 cc. of the clear urine, with 5 cc. of 
of the sodium acetate solution are poured into a beaker 
and heated ; to this the uranium acetate is slowly added 
from the burette. The mixture is constantly stirred 
with a glass rod, which should be applied frequently to 
the cochineal solution. As soon as the green color is 
obtained the process is completed. Read off from the 
burette the amount of uranium acetate solution used. 

Example. Urine in 24 hours equals 1180 cc. Solu- 
tion uranium acetate used equals 24.3 cc. 

.005 X 24.3 cc. 



50 



X 1180=2.86 grams P 2 5 . 



A shorter and approximately accurate method based 
on the above is as follows : To 10 cc. of the urine in a 
test tube add 1 cc. of sodium acetate and 1 cc. of the 
cochineal solution and boil. Add 5% uranium nitrate 
.solution, 1 minim at a time, boiling - after each addition, 
until the mixture turns green. Each minim of the 
uranium solution represents 0.046 gram of P 2 5 in 
1000 cc of urine. Multiply .046 by the number of min- 
ims required to produce the green color, and the product 
will represent the amount in grams of P 2 5 per liter in 
the given sample of urine. 
Estimation of Total Sulphuric Acid. (Estimated as S0 3 ). 
Dissolve 30.5 grams of pure crystallized chloride of barium in 



54 

some distilled water and dilute to 1 liter. Each cc. of this solu- 
tion will equal .01 gram of S0 3 . A dilute solution of sodium 
or magnesium sulphate will also be required. 

Application. 50 cc. of clear urine are poured into a 
beaker, acidified quite strongly with hydrochloric acid, 
and heated over the flame. Let the solution boil for 
about ten minutes, the lamp is then removed and the 
barium chloride is allowed to flow slowly from the bu- 
rette into the beaker and it must continue to flow as 
long as the precipitate is seen to increase. The precipi- 
tate is allowed to subside, then more of the barium 
chloride is added, and this process repeated, until no 
further precipitate is produced. Much time and labor 
will be saved by filtering a few drops of the solution 
now and then, and allowing these to drop into the test 
tube containing some of the dilute sodium or magnesium 
sulphate. As soon as an excess of the barium chloride 
has been added, a precipitate will appear in the test tube. 
Read off from the burette the amount of barium chloride 
used ; each cc. of which will indicate .01 of a gram of 
S0 3 in each cc. of urine, and from this the total amount 
may be calculated. 

Example: Urine 2000 cc. Amount of barium chlor- 
ide solution 25 cc. 

01 y ?*> 

5 P X2000=10 grams SO s . 

Relation of Urinary Constituents in Normal Human Urine. 
There is normally a direct proportion within narrow limits, 
between the solid substances of the urine, which it is desirable 
to keep in mind. 

Relation of Urea to total solids 50% or one-half. 
" " inorganic matter to other solids 30%. 

" " uric acid to urea 2.5% or ^. 

" " nitrogen in urea to total nitrogen 91%. 

" " phosphoric acid to urea 12.5% or y%. 

" " chloride of sodium to urea 40%. 

" " the sulphates to total nitrogen 18%. 

Upon special blanks test specimens of urine quantitatively, 
the number and amount of constituents being unknown. 



55 

IX. 
CHEMICAL EXAMINATION OF URINARY DEPOSITS. 

There is generally a more or less voluminous deposit in the 
urine after standing for 24 hours ; sometimes this deposit is 
formed in the bladder and sometimes it forms after the emis- 
sion of the urine ; it is important to note this fact. Deposits 
or sediments should not be confounded with calculi. The 
former have a pulverulent form while the latter have the 
appearance of grains or granules of greater or less size. 

It is often very easy to determine the nature of the deposit 
by a microscopic examination, but chemical reagents in some 
cases give more precise results. 

With the urine containing sediment, shake thoroughly to 
distribute it, then pour into a centrifugal tube and revolve 
until the sediment is completely precipitated. Note the reac- 
tion of the urine, whether acid or alkaline. Pour off the 
clear supernatant fluid. If the urine is acid pursue the follow- 
ing scheme. 

Acid Urate of Soda. If the deposit is more or less red, 
treat with a little boiling water. If the sediment is dissolved 
it is composed of the acid urate of soda. Caustic potash does 
not dissolve it, hydrochloric acid gives a crystalline precipitate, 
the deposit will give the murexide test. (Page 23). 

Uric Acidi If a crystalline deposit, almost insoluble in 
boiling water, but soluble in caustic potash and giving the 
murexide test, it is uric acid. 

Cystine, (very rare). The deposit is crystalline, insolu- 
ble in caustic potash but soluble in ammonia and hydrochloric 
acid. 

Pus. A white, dense, mucus deposit, viscid and not often 
mixing with the urine ; caustic potash renders it more viscid ; 
the microscope shows the pus corpuscles. This deposit is rare 
in acid urines. 

Blood. Red deposit; the tincture of guaiac with hydrogen 
dioxide gives a blue color. The spectroscope gives the char- 
acteristic lines in the spectrum. The hemin test will^show 
the characteristic crystals. When blood is present the urine 
is generally albuminous. 



56 






C? 



cP9 



Q 







Plate II. 
Various forms of Calcium Carbonate Crystals. (Horse Urine). 



57 



Some deposits not very abundant and having no special 
chemical reaction may be recognized under the microscope. 

Neutral or Alkaline Urine. 

Alkaline Urates. More or less reddish deposits easily solu- 
ble in boiling water and giving the murexide test. 

Triple Phosphate or Calcium Phosphate. A white deposit, 
insoluble in boiling water, soluble in acetic acid, does not give 
the murexide test ; acidified with nitric acid, the molybdate 
of ammonia solution gives a yellowish precipitate in the cold. 

Calcium Oxalate. White deposit insoluble in boiling water, 
also in caustic potash and acetic acid. Soluble in hydrochloric 
acid or in nitric acid ; does not give the murexide test. 

Calcium Carbonate. White deposit insoluble in boiling 
water or in caustic potash. Soluble with effervescence in acetic, 
nitric or hydrochloric acids, does not give murexide reaction. 

Pus, (more frequent in alkaline and albuminous urine). 
A white, dense, mucus-like deposit, not mixing readily with 
the rest of the urine. Caustic potash renders it more viscid, 
the microscope shows pus globules. 

Blood. Determined by the guaiacum test, spectroscope, 
microscope, or hemin test. 

From the pathologic point of view the presence of urin- 
ary sediments generally indicate an alteration of the secretions. 



A 








1 III 


' I 






i 1 ! M i I 1 i M 


B 










Ill 












RED: . ORANGE YELL W GfiEEN BLUE VIOLET; 


c 


' 






! 






1 







Fig. 16. 

A. — Spectrum of Oxyhemoglobin. B. — Spectrum of Reduced Hemoglobin. 
C— Spectrum of Urobilin. 



58 




Fig. 17. Uric Acid Crystals. 



Microscopical Examination of Unine. If an immed- 
iate examination is desired the centrifuge may be used to 
cause the sediment to separate from the urine ; otherwise the 
urine is set aside for some hours in a cool place when the sedi- 
ment gradually settles to 
the bottom. The super- 
natant liquid is poured off 
and by means of a pipette, 
camel's hair brush- or a 
wire loop a small portion 
of the deposit is trans- 
ferred to a slide and ex- 
amined. 

The deposits may be 
divided into unorganized 
and organized sediments. 
Unorganised Sediments. In unorganized sediments two con- 
ditions may be considered : 1st, the urine is acid. 2nd, the 
urine is alkaline. 

In acid urine look first for 
crystals of uric acid diverse in 
form and reddish brown in color. 
(Fig. 17). Second, crystals of acid 
urate of soda, yellowish or red. 
The crystals are badly formed. 
Third, crystals of oxalate of lime 
also found in alkaline urine. (Fig. 
18). Fourth, crystals of hippuric 
acid. Fifth, crystals of calcium 
sulphate. (4 and 5 are met with 
rarely in acid urine). Sixth, cal- 
cium phosphate. Make sure of the identification by reference 
to charts. 

In alkaline urine look first for ammonio-magnesium phos- 
phates, (Fig. 19), (triple phosphates). Second, bicalcium 
phosphate crystals. Third, tricalcium or amorphous phos- 
phate crystals. Fourth, crystals of sulphate of lime. Fifth, 




Fig. 18. Calcium Oxalate. 



59 




Fig. 19. Triple Phosphates. 

crystals of oxalate of lime, (also found in acid urine), Sixth, 
crystals of urate of ammonium in the form of yellow colored 
spheres. 

Crystals of cystin, leucin and tyrosin are sometimes en- 
countered in acid or alkaline urines. 

Organised Sediments. These are brought more plainly into 
view if the preparations are stained, although this is not uni- 
versally practised. Organized sediments may be divided into 
(a) histologic elements, (b) microbic elements. 

Of the histologic elements there 
are frequently encountered, 1st, epi- 
thelial cells from the bladder, from the 
vagina and from the uterer, their pres- 
ence has no special significance. If, 
however, they are present in consider- 
able quantity, a lesion of these parts 
may be suspected. 2nd, cells from the 
pelvis of the kidney, generally an indi- 
cation of renal affection. 3rd, cancer 
cells, which have a special significance. 
4th, hyaline or granular casts, seen more easily in stained 
preparations ; they are encountered frequently in albuminous 
urine ; their presence indicates a mild form of nephritis. 5th, 
epithelial, hemorrhagic or wax casts, of which the hemorrhagic 
are the most frequent, they are markedly colored and easily 
recognized on account of the hemoglobin they contain ; they 
contain fine granulations which are not blood corpuscles but 




Fig. 20. 
Micrococcus Ureae. 



60 




possible fragments 
of them, they indi- 
cate a severe form 
of nephritis. 6th, 
cylindroid ele- 
ments drawn out 
in an irregular 
and somewhat 
ribbon-like form ; 
they are the pro- 
duct of the secre- 
In the human race they may 



Normal. 



Fig. 21. Red blood corpuscles. 
B. Decolorized. 

tion of the urinary epithelium, 
be found in cases of scarlatina and generally in certain forms 
of nephritis. In examining for casts it is desirable to examine 
the urine immediately after emission, as they generally disin- 
tegrate rapidly and disappear. 7th, blood corpuscles more 
or less crenated but easily recognizable by their yellowish 
tint ; the presence of blood corpuscles in the urine indicate a 
hemorrhage either in the bladder or in the kidney. 8th, pus 
corpuscles which 
may have crenated 
borders, granular 
contents and ap- 
pear quite refrac- 
tive, a drop of 
dilute acetic acid 
will render the 
nuclei visible. Pa- 
thologically the 
pus corpuscles in- 
dicate a suppura- 
tion of some por- 
tion of the urinary 
tract. In the ma- 
jority of cases it is 
impossible to say 
if the pus comes 
from the bladder 

or kidney. In cases of cystitis of the neck of the bladder, 
the last portion of the urine passed contains no pus, while if 




Fig. 22. Red blood corpuscles crenated. 










E 




1 a "" 




H 



# /it? 









£7 






1 






<*><# 



<»© 









*>a>*® * 









*> 






4L\ 



^ a 



/,< jyh A 

6fr r¥ 





AT 4SF 

49 ^ JTtifr " 



%}& 




PLATE III. 
Urinary Sediment. A. Uric Acid. B. Ammonium Urate. C. Acid Sodium 
Urate. D. Urea Nitrate. E. (t) Leucin and (2) Tyrosin. F. Cystin. G. Ammonio- 
Magnesium or triple phosphate. H. Calcium Phosphate. I. Calcium Oxalate. J. 
Blood Corpuscles. K. Pus and Mucus. L, Hemin Crystals. M. (t) Hyaline Casts, 
(2) Granular Casts. N. Epithelial Casts and Cells. 0. (l) Waxy Casts ; (2) Casts 
with Blood Corpuscles ; (3) Casts with Fat Globules. (After Simon). 



62 



this trouble be in the kidney this last portion will contain 
pus. This fact is of some use in diagnosing cystitis of the 
neck of the bladder. 9th, spermatozoa may be present but 
are easily recognized by their elongated form. 10th, mucus 
may be frequently present, but has no very great pathologic 
significance. 

Urinary Casts. Although probably observed earlier, 
Henle is credited in 1842, with first carefully describingthe casts 
moulded in the tubules of the kidneys. Among earlier views, 
casts were considered as being composed of coagulated fibrin ; 
as products of the secretion of the epithelium of the tubules ; 
as transformed or disintegrated epithelial cells and as prod- 
ucts from the blood. A view quite commonly accepted is that 
casts are the products of the coagulation of albuminous mater- 
ial. The fact that the presence of casts in the urine is usually 
accompanied by the presence of albumin lends force to this 
view ; for the more abundant the albumin, the more likelihood 
is there of finding casts. From this standpoint, then, casts 
may be regarded as albuminous exudates from the blood, with 
the addition of transformed or destroyed epithelium. In 
nearly all cases where casts are present, some albumin is 
found. Occasionally they are sometimes described as being 
present without the appearance of albumin. This condition 
is looked upon with some doubt ; for it is commonly believed 
that albumin exists but in an amount too slight to be detected 
by the ordinary chemical tests. 

The presence of casts in the 
urine is of much diagnostic im- 
portance ; if found in any quantity, 
they indicate nephritis particu- 
larly if albumin is also present in 
any amount. It is claimed by 
some that merely hyperemia of 
the kidney will cause the appear- 
ance of casts, in the urine, and 
that they may sometimes be found 
when the kidneys are perfectly 
intact. Mitchell states that two 

or three hyaline or one or two 

,« , , Fig. 23. a. Showing forma- 

small granular casts may be found tion of hyaline cast in tubule b 

Hyaline cast with granular deposit, 
c. Granular cast. 




63 



in one of every three specimens of the twenty-four hour's urine 
examined (human). Casts have been described in cases of 
g-astro-intestinal catarrh, in jaundice, in acute and chronic 
anemia, as well as in nervous affections of different kinds, 
without accompanying- inflammation of the kidneys. There 
are many, however, who hold that casts are always the pro- 
ducts of an inflammatory process, and are, therefore, indicative 
of renal inflammation. Other evidences or symptoms should 
also be taken into account. 

Occasion- 
ally it is dif- 
ficult to find 
casts even 
when they 
are known to 
exist. Alka- 
line urine has 
a tendency to 
dissolve them 
At times 
they will not 
settle for 
hours. Usu- 
ally if al- 
lowed to 
stand six 
hours, the 
casts will set- 
tle if they are 
present. The 
cen trif u g e 

will bring- them down in a few minutes. A low power of the 
microscope (150-200 diameters) may be used in the search for 
casts and will enable the observer to pass over the field quite 
rapidly. To identify the cast and its structure, a power of 
400-500 diameters is required. More than one specimen must 
always be examined before giving a positive statement as to 
the presence or absence of casts. False or pseudo-casts have 
been described ; these are believed to be accidental formations, 
while true casts in g-eneral indicate nephritis. 




Fig. 24. a. 
blood cells, b. 
thelial cells, d. 



Blood cast and hyaline cast carrying 
Pus cast. c. Hyaline cast carrying epi- 
Epithelial casts. (Greene). 



64 



True casts may appear in three different sizes according 
to the portion of the tubule in which they are formed. The 
smallest size originate in the narrow tubule. The next in 
size come from the convoluted tubule of the second order (no 
casts from the convoluted tubule of the first order, i. e., that 
portion of the tubule nearest the glomerules, appear in the 
urine, because they are too large to pass through the narrow 
tubules ). The third and largest are those developing in the 
straight collecting tubules. It is believed a prognostic value 
may be attached to the size of the casts as well as to their 
number. Casts from the narrow tubules indicate a mild 

attack ; from 
the convo- 
luted tubules 
a severe 
form, especi- 
ally in the 
cortex of the 
kidney ; from 
the collect- 
ing tubules, 
with the 
ot h er forms 
also present, 
a serious con- 
d i t i o n of 
general renal 
inflammation 
or unfavor- 
able prognos- 
is. The iden- 
tity of casts 
may generally be determined by their uniform width. They 
are usually longer than they are broad, and have one well- 
rounded extremity and well defined borders. 

True casts are of six varieties ; hyaline, epithelial, blood, 
granular, fatty and waxy casts. In a general way the first 
three varieties are found in an acute form of nephritis. Dur- 
ing the first few weeks of the inflammation the last three 
varieties are not encountered. If the acute condition passes 




Fig. 25. Granular Casts. (Green). 



65 



into a subacute, then the granular variety appears, at first in 
small number, then in larger, with, usually, a considerable 
number of the hyaline and epithelial casts. Fatty and waxy 
casts are always secondary products, and as a rule not found 
until a nephritis has existed for some time. 

Hyaline casts are 
colorless, pale, more 
or less transparent 
formations soluble in 
acetic acid. They 
are of variable size 
and generally diffi- 
cult to detect on ac- 
count of their appar- 
ent 1 y structureless 
character. At times 
a slight granulation 
may be seen imbed- 
ded in or adhering 
to their matrix and 
occasionally acci- 
dental attachments 
of pus or fat globules 
in small numbers. 

Epithelial casts have a hyaline matrix more or less con- 
cealed by epithelial cells. The presence of these casts is in- 
dicative of an acute process. 

Blood casts consist of 
the hyaline matrix with 
blood corpuscles imbedded 
in or adhering to the ma- 
trix. Pus casts are rare, 
but when present the pus 
corpuscles adhere to the 
matrix. Blood casts are 
indicative of a hemorrh- 
age into the tubules and 
of an acute hemorrhagic 
process. Hyaline and epithelial casts are usually associated 
with them. 




Fig. 26. 
casts, d. 



a. Fatty casts, b. and c. Blood 
Free fatty Molecules. (Roberts). 




Fig. 27. Fatty Casts and Fat Droplets. 



66 



Graular casts usually have well defined boundaries with 
granular matter imbedded in or adhering- to the matrix. They 
may be finely or coarsely granular, the latter having a more 
serious significance. Granular casts are due to a disintegra- 
tion of the renal epithelium. Their degree of refraction is 
changeable ; sometimes they appear yellowish, at other times 
colorless. 

Fatty casts have a hyaline matrix containing a number of 
small, glistening fat globules and granules. Some free fat is 
also usually found in the field. As fatty casts are secondary 
products of epithelial and granular casts, the diagnosis of a 
chronic process is justifiable. • The hyaline matrix is char- 
acteristic of the different varieties of casts that have been 
mentioned up to this point- 
Waxy casts differ in 
chemical composition from 
those previously mention- 
ed. They are character- 
ized by wavy contours ; a 
high refracting power ; a 
more or less yellowish col- 
or and quite a high degree 
of brittleness. They are 
slowly, if at all, attacked 
by acetic acid. Their 
presence signifies waxy 
degeneration of the kid- 
ney. Hyaline casts may 

sometimes have a superficial resemblance to waxy casts, but 
they never have the same high refraction as the latter. 

Cast-like formations are composed of various elements 
having a form somewhat similar to casts but lacking the 
matrix soluble in acetic acid. Amorphous urates often simu- 
late granular casts in form. Bacteria are often grouped in a 
manner similar to the form of a cast, but a close inspection 
shows an irregular outline, and usually a number of groupings 
not in cast form. Granular detritus and hematoidin may also 
assume the form of casts ; likewise epithelial cells, blood cor- 
puscles and fibrin in renal hemorrhages may also assume the 
form of casts. Acetic acid is said to be a reliable reagent for 




Fig. 28. Waxy Casts. 



67 



differentiating between true and false casts. This reagent 
dissolves the matrix of the true casts but does not act upon the 
cast-like formations. 

Cylindroids appear like hyaline 
casts, but are large and band-like. 
Their breadth is uniform and they 
often contain crystals, epithelial 
cells and corpuscles. They are sol- 
uble in acetic acid and are of renal 
origin. No especial significance is 
attached to them. 

Mucous cylinders, sometimes 
called cylindroids, are usually not of 
uniform breadth and seldom contain 
morphologic constituents and are in- 
soluble in acetic acid. They are 
usually found in any urine containing an abundance of mucus 
and are of no special significance. 




Fig. 29. Cylindroids. 
a, b, Cast-like forms; c, Fil- 
amentous forms. (Ogden). 



68 



Form used in examination of Horse Urine. 



Owner Address 



No. Species.. 



Age 



..Sex 







Normal (Horse). 


Sample 


Amount in 24 hours 


3000-4000 c. c. 




Specific Gravity 


1025-1050 




Reaction 


Alkaline 




Color 


Yellowish brown 




Translucency 


Turbid 




Consistency 


Viscid 




Total Solids 


50-120 parts per 1000 




Chlorides 


8-14 




Sulphates 


2-3 


. 


Phosphates 


.05.2 




Urea 


20-40 




Uric Acid 


Trace 




Hippuric Acid 


4-8 parts per 1000 




Indican 


.1-.2 " 




ABNORMAL CONSTITUENTS. 




Albumin 






Sugar 






Bile 






Hemoglobin 






MICROSCOPIC EXAMINATION. 




Epithelial Cells 






Leucocytes 






Blood 






Casts 






Spermatozoa 






Micro-organisms 






Calcium Carbonate 






Calcium Oxalate 






Triple Phosphates 







69 



Form used in Examination of Human Urine. 



Name. 



Address. 



Date. 
Amount in 24 hours. 
Specific gravity 
Reaction 
Color 
Translucency 

Solids 

Chlorides 

Phosphates 

Sulphates 

Urea 

Uric Acid 
Indicau 



Normal. Sample. 

1000-1500 cc 
1015-1025 
Acid 

Light golden 
Clear 

34-50 parts per 1000 
50-75 parts per 24 hours 
6-9 parts per 1000 as NaCl 
10-15 parts per 24 hours 
1.5-2 5 parts per 1000 as P 2 5 
2.-3.5 parts per 24 hours 
1.5^3 parts per 1000 as SO 3 
2.5-4 parts per 24 hours 
14-22 parts per 1000 
22-33 parts per 24 hours 
0.25-0.40 parts per 1000 
0.40-0.60 parts per 24 hours 



Albumin 
Sugar 
Bile 
Hemoglobin 



MICROSCOPIC EXAMINATION 



Epithelial cells 

Leucocytes 

Blood 

Casts 

Uric Acid 

Oxalates 

Triple Phosphates 

Urates 



THE PATHOLOGY AND DIFFERENTIAL DIAGNOSIS 
OF INFECTIOUS DISEASES OF ANIMALS. By Veranus Alva 
Moore, B. S., M. D., Professor of Comparative Pathology, Bacteriology 
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Elementary Exercises in Materia Medica 
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A laboratory manual treating of the more commonly used inorganic 
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A full list of drugs with their doses for the different domestic animals 
based upon the eighth (1905) revision of the U. S. Pharmacopoeia. 
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Second Edition. Revised and Enlarged. Red Leather Flexible 
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A laboratory manual. Part I., Chemical Physiology. Part II., Ex- 
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Second Edition. Revised and Enlarged. $1.50 net. 

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n Journal of Ap. Microscopy and Laboratory Methods." 



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Veterinary Medicine 

By Professor James Law, F. R. C. V. S. 

Director N. Y. S. Veterinary College Ithaca, N. Y. 

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Examination of the Urine of the Horse and Man. 

By Pierre A. Fish, D. So, D. V. M. 

Simple directions are given for performing the tests necessary to d< 
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GENERAL SURGERY 

BY DR. MED. EUGEN FROHNER 

Professor in the Royal Veterinary High School in Berlin. Authoriz- 
ed Translation from The Third Revised Edition, by D. Hammond Udall, 
B. S. A., D. V. M., Associate Professor of Surgery and Obstetrics, Ohio 
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The translation of Frohners General Surgery has been undertaken to 
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SL° 37 1906 



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